重组质粒DNA的规模化生产工艺及质量鉴定研究
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摘要
基因治疗和基因免疫是目前生物医药领域研究的热点之一,其中以表达目的基因重组质粒为基础的非病毒基因转移策略具有安全性好等突出优点,更加受到研究者的青睐。目前实验室制备重组质粒DNA不仅技术较为成熟,而且有众多的商品化试剂盒可供选用,但这些试剂盒不仅价格昂贵,而且不能满足大规模制备质粒DNA的需要。大规模制备既符合相关质量要求,价格又便宜的重组质粒DNA,特别是动物用重组质粒DNA,仍是制约核酸疫苗和基因治疗研究的瓶颈。
目前,大规模制备质粒DNA的方法主要有阴离子交换、分子排阻、疏水色谱等层析方法,这些技术不仅需要用苯酚等有毒化学物质和具有潜在病原污染的动物源RNA酶对质粒DNA进行前处理,而且具有耗时、产量低和成本昂贵等缺点。为了解决动物基因免疫或基因治疗用重组质粒DNA的规模化生产工艺,本研究在筛选出具有价格优势的重组菌高密度发酵培养基配方的基础上,研究了不同重组质粒DNA转化大肠杆菌的发酵培养条件,发酵培养过程中宿主菌生长动态与质粒DNA复制的关系,并以150L发酵罐为标准,研究了不同发酵培养条件下重组质粒DNA的产量。结果表明,DH5α和JM109重组菌在筛选的发酵培养基中的生长动态及单位湿菌重比在进口试剂配制的标准LB培养基中要高,在生长对数中后期对重组菌进行42℃诱导,可显著提高质粒产量,其中JM109宿主菌较DH5α宿主菌的质粒产量约高20%。
规模化生产重组质粒DNA的另一重要环节是重组菌的破碎处理和质粒DNA的纯化。目前常用的细菌破碎处理方法包括热裂解和碱溶解法,前者需要专门设计的仪器设备,裂解条件不易控制,后者不需专门设备,但生产成本较高。本研究对标准的重组大肠杆菌碱溶解法进行了改进,使其在不影响质粒产量和质量的前提下,制备成本明显下降。进而在公开发表的十六烷基三甲基溴化铵(CTAB)
Gene therapy and gene immunization are hot research spots in current biomedical field. Among the gene transfer strategies under investigation, the non-viral gene transfer using the recombinant plasmids expressing the gene of interest has the overt advantages of good safety and ease of preparation over viral-mediated vectors. In terms of plasmid DNA preparation, currently-available commercial kits have the disadvantages of small-scale preparation, high cost and requirement of pretreatment using toxic chemicals, which are unsuitable for large-scale preparation of recombinant plasmids for animal use. As to other methods for large-scale production of recombinant plasmids such as anion-exchange chromatography, size-exclusion chromatography and hydrophobic chromatography, pretreatments using toxic chemicals such as phenol and chloroform and animal-derived RNase A are required, as well as time-consuming and high cost. To establish a suitable method for large-scale preparation of plasmid DNA for animal gene immunization/therapy, cost-effective culture media and the fermentation conditions were established for high-density fermentation of recombinant E.coli. Furthermore, the relationship between the plasmid replication and host bacterial growth was investigated in a 150-liter fermentator. The results showed that the growth state and the wet weight of recombinant DH5αand JM109 E.coli in the modified medium were better than that in standard LB medium. By shifting the culture temperature to 42℃after mid-log growth phase, the plasmid yield could be greatly improved.
Other bottlenecks in the industrial production of plasmid DNA include disruption
目前,大规模制备质粒DNA的方法主要有阴离子交换、分子排阻、疏水色谱等层析方法,这些技术不仅需要用苯酚等有毒化学物质和具有潜在病原污染的动物源RNA酶对质粒DNA进行前处理,而且具有耗时、产量低和成本昂贵等缺点。为了解决动物基因免疫或基因治疗用重组质粒DNA的规模化生产工艺,本研究在筛选出具有价格优势的重组菌高密度发酵培养基配方的基础上,研究了不同重组质粒DNA转化大肠杆菌的发酵培养条件,发酵培养过程中宿主菌生长动态与质粒DNA复制的关系,并以150L发酵罐为标准,研究了不同发酵培养条件下重组质粒DNA的产量。结果表明,DH5α和JM109重组菌在筛选的发酵培养基中的生长动态及单位湿菌重比在进口试剂配制的标准LB培养基中要高,在生长对数中后期对重组菌进行42℃诱导,可显著提高质粒产量,其中JM109宿主菌较DH5α宿主菌的质粒产量约高20%。
规模化生产重组质粒DNA的另一重要环节是重组菌的破碎处理和质粒DNA的纯化。目前常用的细菌破碎处理方法包括热裂解和碱溶解法,前者需要专门设计的仪器设备,裂解条件不易控制,后者不需专门设备,但生产成本较高。本研究对标准的重组大肠杆菌碱溶解法进行了改进,使其在不影响质粒产量和质量的前提下,制备成本明显下降。进而在公开发表的十六烷基三甲基溴化铵(CTAB)
Gene therapy and gene immunization are hot research spots in current biomedical field. Among the gene transfer strategies under investigation, the non-viral gene transfer using the recombinant plasmids expressing the gene of interest has the overt advantages of good safety and ease of preparation over viral-mediated vectors. In terms of plasmid DNA preparation, currently-available commercial kits have the disadvantages of small-scale preparation, high cost and requirement of pretreatment using toxic chemicals, which are unsuitable for large-scale preparation of recombinant plasmids for animal use. As to other methods for large-scale production of recombinant plasmids such as anion-exchange chromatography, size-exclusion chromatography and hydrophobic chromatography, pretreatments using toxic chemicals such as phenol and chloroform and animal-derived RNase A are required, as well as time-consuming and high cost. To establish a suitable method for large-scale preparation of plasmid DNA for animal gene immunization/therapy, cost-effective culture media and the fermentation conditions were established for high-density fermentation of recombinant E.coli. Furthermore, the relationship between the plasmid replication and host bacterial growth was investigated in a 150-liter fermentator. The results showed that the growth state and the wet weight of recombinant DH5αand JM109 E.coli in the modified medium were better than that in standard LB medium. By shifting the culture temperature to 42℃after mid-log growth phase, the plasmid yield could be greatly improved.
Other bottlenecks in the industrial production of plasmid DNA include disruption
引文
1. Prazeres DM. Purification of plasmids for gene therapy and DNA vaccination. Biotechnol Annu Rev, 2001, 7: 1-30
2. 吴乃虎. 基因工程原理. 科学出版社(第二版), 2001
3. Carole L. Bassett and Sidney R. Kushner. Exonucelases I, III and V are required for stability of ColE1-Related Plasmids in Escherichia coli. Journarl of Bacteriology, 1984, 157 (2): 661-664
4. Joachim Stadler, Raf Lemmens and Tomas Nyhammar. Plasmid DNA purification. The Journal of Gene Medicine, 2004, 6 (1): S54-66
5. 张玉静 主编. 分子遗传学. 科学出版社, 2002
6. J.萨姆布鲁克, D.W 拉塞尔. 分子克隆实验指南第三版. 科学出版社, 2003
7. Wang Z, Yuan Z, Hengge UR. Processing of plasmid DNA with ColE1-like replication origin. Plasmid, 2004, 51 (3): 149-61
8. Chambers S, Barstow D, Minton N. The pMTL nic-cloning vectors I Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene, 1988, 68 (1): 139-49
9. Yavachev L, Ivanov I. What does the homology between E. coli tRNAs and RNAs controlling ColE1 plasmid replication means?. J. Theor. Biol, 1988, 131 (2): 235-241
10. Wang,Z, Le,G, Shi Y, et al. A model for regulation of ColE1-like plasmid replication by uncharged tRNAs in amino acid-starved Escherichia coli cells. Plasmid, 2002c, 47 (2): 69-78
11. Minton NP. Improved plasmid vectors for the isolation of translational lac gene fusions. Gene, 1984, 31 (1-3): 269-73
12. N.A. Horn, J.A. Meek, G. Budahazi, M. Cancer gene therapy using plasmid DNA: purificationof DNA for human clinical trials. Human Gene Therapy, 1995, 6 (5): 565-573
13. N.C. Wan, J.C. Goodrick. Method for increasing plasmid yield. United States Patent#5, 1996, 487, 986
14. R. Lahijani, G. Hulley, G. Soriano, N.A. Horn, et al. High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature-controllable point mutation. Hum Gene Ther, 1996, 7 (16): 1971-80
15. Uhlin B, Nordstr?m K. A runaway-replication mutant of plasmid R1drd-19: temperature-dependent loss of copy number control. Mol Gen Genet, 1978, 165 (2): 167-79
16. Remaut E, Tsao H, Fiers W. Improved plasmid vectors with a thermo-inducible expression and temperature-regulated runaway replication. Gene, 1983, 22 (1): 103-13
17. Ansorge MB, Kula MR. Investigating expression systems for the stable large-scale production of recombinant leucine-dehydrogenase from Bacillus cereus in Escherichia coli. Appl Microbiol Biotechnol, 2000, 53 (6): 668-73
18. Chao Y, Chern J, Wen C. Coupling the T7 A1 promoter to the runaway-replication vector as an efficient method for stringent control and high-level expression of lacZ. Biotechnol Prog, 2001, 17 (1): 203-7
19. Kristala Jones Prather, Sangeetha Sagar, Jason Murphy, et al. Industrial scale production of plasmid DNA for vaccine and gene therapy: plamsid design, production, and purification. Enzyme and Microbial Technology, 2003, 33 (7): 865-883
20. Smith H. Regulatory considerations for nucleic acid vaccines. Vaccine, 1994, 12 (16): 1515-9
21. Gurunathan S, Klinman D, Seder R. DNA vaccines: immunology, application, and optimization. Annu Rev Immunol, 2000, 18: 927-74
22. Davis M, Huang E. Transfer and expression of plasmids containing human cytomegalovirus immediate-early gene promoter-enhancer sequences in eukaryotic and prokaryotic cells. Biotechnol Appl Biochem, 1988, 10 (1): 6-12
23. Chapman B, Thayer R, Vincent K, et al. Effect of intron A from human cytomegalovirus (Towne) immediate-early gene on heterologous expression in mammalian cells. Nucleic AcidsResearch, 1991, 19 (14): 3979-86
24. Verfailliea T, Cox B.M. Goddeeris. Immunostimulatory capacity of DNA vaccine vectors in porcine PBMC: a specific role for CpG-motifs. Veterinary Immunology and Immunopathology, 2005, 103 (1-2): 141-151
25. Klinman D M. CpG DNA as a vaccine adjuvant. Expert Rev Vaccines, 2003, 2 (2): 305-15
26. FDA. Points to consider on plasmid DNA vaccines for preventive infectious diseases. 1996, Docket No. 96N-0400
27. Robertson J, Griffiths E. WHO guidelines for assuring the quality of DNA vaccines. Biologicals, 1998, 26 (3): 205-12
28. Falk LA, Ball LK. Current status and future trends in vaccine regulation--USA. Vaccine, 2001, 19 (13-14): 1567-72
29. Cichutek K. DNA vaccines: development, standardization and regulation. Intervirology, 2000, 43 (4-6): 331-8
30. Robertson JS, Cichutek K. European union guidance on the quality, safety and efficacy of DNA vaccines and regulatory requirements. Dev Biol, 2000, 104: 53-6
31. Smith HA. Regulatory considerations for nucleic acid vaccines. Vaccine, 1994, 12 (16): 1515-9
32. Smith HA. Regulation and review of DNA vaccine products. Dev Biol, 2000, 104: 57-62
33. W Chen, C Graham and RB Ciccarelli. Automated fed-batch fermentation with feed-back controls based on dissolved oxygen (DO) and pH for production of DNA vaccines. Journal of industrial Microbiology and Biotechnology, 1997, 18 (1): 43-48
34. 赵 霞, 叶 勤, 俞俊棠. 影响重组工程菌发酵产率的因素. 工业微生物, 1995,25(2): 26-32
35. Erik Remaut, Hsu Tsao and Walter Fiers. Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication. Gene, 1983, 22 (1): 103-113
36. Robin-Ann Klotsky and Ira Schwartz. Measurement of cat expression from growth-rate-regulated promoters employing β-lactamase activity as an indicator of plasmid copy number. Gene, 1989, 55 (1): 141-146
37. 邱英华, 沈 益, 王玉海, 等. Cecropin X 发酵过程中工程菌质粒稳定性的研究. 微生物学报, 2004, 44 (3): 390-391
38. Jos ML Secgers, Cristian M Franke, Rense K, et al. Use of continuous culture for the selection of plasmids with improved segregational stability. Plasmid, 1995, 33 (1): 71-77
39. Nordstrom K, Molin S, Aagaard-Hansen H. Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid, 1980, 4 (2): 215-27
40. Engberg B and Nordstrom, K. J. Plasmid stability in E.coli. J. Bacteriol, 1975, 179: 123
41. Futchjer AB, Cox BS. Copy number and the stability of 2-micron circle-based artificial plasmids of Saccharomyces cerevisiae. J.Bacteriol, 1984, 157 (1): 283-90
42. Caulcott CA, Dunn A, Robertson HA, et al. Investigation of the effect of growth environment on the stability of low-copy-number plasmids in Escherichia coli. J Gen Microbiol, 1987, 133 (7): 1881-9
43. 杨海燕, 方呈祥, 张珞珍, 等. 不同质粒在大肠杆菌宿主细胞中的不稳定性研究. 河南农业大学学报, 1999, 33 (2): 190-193
44. Chang LL, Hwang LY and Hwang CF, et al. Study of high density Escherichia coli fermentation for production of porcine somatotropin protein. Ann N Y Acad Sci, 1991, 27 (5): 259-72
45. Ross H. Durland, Eric M. Eastman. Manufacturing and quality control of plasmid-based gene expression systems. Advanced Drug Delivery Reviews, 1998, 2 (30): 33-41
46. Takapa 商品目录, 2004-2005
47. 许乃用. 稳定质粒拷贝数提高重组蛋白质的产量. 工业微生物, 2002, 32 (2): 58
48. 姜泊, 张亚力. 分子生物学常用试验方法. 北京: 人民军医出版社, 1996
49. 卢圣栋. 现代分子生物学实验技术. 北京: 高等教育出版社, 1999, 54
50. Lahijani R, Hulley G, Soriano G, et al. High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature controllable point mutation. Human Gene Therapy, 1996, 7 (16): 1971-80
51. Schmidt T, Friehs K, Schleef M, et al. In-process analysis of plasmid copy number for fermentation control. Pacesetter, 2001, 5 (1): 4-6
52. Korz DJ, Rinas U, Hellmuth K, et al. Simple fed-batch technique for high cell densitycultivation of Escherichia coli. J Biotechnol, 1995, 39 (1): 59-65
53. O’Kennedy RD, Ward JM, Keshavarz-Moore E. Effects of fermentation strategy on the characteristics of plasmid DNA production. Biotechnol Appl Biochem, 2003, 37 (1): 83-90
54. Ronan D. O’Kennedy, John M. Ward and Eli Keshavarz-Moore. Effects of fermentation stategy on the characteristics of plasmid DNA production. Biotechol. Appl. Biochem, 2003, 37: 83-90
55. R.D. O’Kennedy, C.Baldwin, E.Keshavarz-Moore. Effects of growth medium selection on plasmid DNA production and initial processing steps. Journal of Biotechnology, 2000, 76 (2-3): 175-183
56. 俞俊棠, 唐孝宣. 生物工艺学. 上海: 华东理工大学出版社, 2000: 211
57. Nakasu S and Tomizawa J. Structure of the ColE1 DNA molecule before segregation to daughter molecules. Proc Natl Acad Sci U S A, 1992, 89 (21): 10139-43
58. D.J. Rorz, U. Rinas, K. Hellmuth, et al. Simple fed-batch technique for high density cultivation of Escherichia coli. Journal of Biotechnology, 1995, 39 (1): 59-65
59. Lahijani R, Hulley G, Soriano G, et al. High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature-controllable point mutation. Hum Gene Ther, 1996, 7 (16): 1971-80
60. Reinikainen P, K Korpela, V Nissine, et al. Escherichia coli plasmid production in fermentator. Biotechnol Bioeng, 1989, 33: 386-393
61. QIAGEN Plasmid Purification Handbook, Second Edition, August 2003
62. Michel Thiry and Doriano Cingotanl. Optimizing scale-up fermentation processes. Trends in Biotechnology, 2002, 20 (3): 103-105
63. Lee SY. High cell-density culture of Escherichia coli Reviews. Trends Biotechnol, 1996, 14 (3): 98-105
64. Docket NO.96N-0400 Points to Consider on Plasmid DNA Vaccines for Preventive Infectious Disease Indications
65. Ellenberg SS. Safety considerations for new vaccine development. Pharmacoepidemiol Drug Saf, 2001, 10 (5): 411-5
66. D.M.F. Prazeres, G.N.M.Ferreira. Design of flowsheets for the recovery and purification of plasmids for gene therapy and DNA vaccination. Chemical Engineering and Processing, 2003 , 4 (43): 615-630
67. D.Petrides. Bioprocess design in: R.G. Harrison. P.W. Todd, S.R.Rudge, D. Petrides (eds). Biseparations Scinece and Engineering, Oxford University Press,Oxford, UK, 2003, pp. 319-372
68. 郭利平, 章佩芬, 唐青涛. 基因治疗和 DNA 疫苗质粒 DNA 的下游加工技术. 华西药学杂志, 2002, 17 (2): 159-162
69. Zhijun Wang, Guowei Le, Yonghui Shi, et al. Studies on recovery plasmid DNA from Echerichia coli by heat treatment. Process Biochemistry, 2002, 38: 199-206
70. Birnboim HC, Doly J. rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res, 1979, 7 (6): 1513-23
71. Jon R. Sayers, Debbie Evans and James B. Thomson. Identification and Eradication of a Denatured DNA Isolated during Alkaline Lysis-Based Plasmid Purification Procedures. Analytical Biochemistry, 1996, 241 (2): 186-189
72. Jun Song, Christian Geltinger and Kailai Sun, et al. Direct Lysis Method for the Rapaid Preparation of Plasmid DNA. Analytical Biochemistry, 1999, 271 (1): 89-91
73. Wen-Chi Tseng, Fu-lyn Ho, Tsuei-Yun Fang, et al. Effect of alcohol on purification of plasmid DNA using ion-exchange membrane. Journal of Membrane Scinece, 2004, 233: 161-167
74. Ferreira GNM. A comparison of gel filtration chromatographic supports for plasmid purification. Biotechnol Techn, 1997, 11: 417-420
75. Diogo MM, Queiroz JA, Monteiro GA, et al. Separation and analysis of denatured plasmid forms using hydrophobic interaction chromatography. Anal Biochem, 1999, 275 (1): 122-124
76. Matteo D. Costioli, lgor Fisch, Frank Hilbrig, et al. DNA purification by Triple-Helix Affinity Precipitation. Biotechnology and Bioengineering, 2003, 81 (5): 535-545
77. Alex Eon-Duval, Rober H. MacDuff, Carol A, et al. Removal of RNA impurities by tangential flow filtration in an RNase-free plasmid DNA purification process. Analytical Biochemistry,2003, 316 (1): 66-73
78. Dan V. Mourich, Michael W. Munks, Jason C. Murphy, et al. Spermine compaction is an efficient and ecomomical method of producing vaccination-grade DNA. Journal of Immunological Methods, 2003, 274 (1-2): 257-264
79. Russel J.Lander, Michael A. Winters, Francis J. Meacle, et al. Fractional Precipitation of Plasmid DNA from Lysate by CTAB. Biotechnology and Bioengineering, 2002, 79 (7): 777-784
80. Per-Olof Wahlund, Per-Erik Gustavsson, Vladimir A. Izumrudov, et al. Polyelectrolyte complexes as a tool for purification of plasmid DNA Background and development. Journal of Chromatograohy B, 2004 , 807 (1): 121-127
81. Raf Lemmens, Urban Olsson, Tomas Nyhammar, et al. Supercoiled plasmid DNA: selective purification by thiophilic/aromatic adsorption. Journal of Chromatography B, 2003, 784 (2): 291-300
82. Sandhya R. Shenoy, Mohammad Khalid, Anshu Gupta, et al. Purification of an Enterobacter aerogenes Plasmid DNA using MnCl2 as compaction agent. Analytical Biochemistry, 2003, 321 (2): 256-258
83. B. Atkinson, F. Mavituna (Eds). Biochemical Engineering and Biotechnology Handbook, Macmillan, New York, USA, 1991
84. Sofer, G, and Hagel, L. Handbook of Process Chromatography: A Guide to Optimization, scale-up and Validation (1st edn), Academic Press
85. G.D.Cooke, R.M.Cranenburgh, J.A.J. Hanak, et al. Purification of essentially RNA free plasmid DNA using a modified Escherichia coli host strain expressing ribonuclease A. Journal of Biotechnology, 2001, 85 (3): 297-304
86. David W.Kahn, Michelle D. Butler, Darien L. Cohen, et al. Purification of Plasmid DNA by Tangential Flow Filtration. Biotechnology and Bioengineering, 2000, 69 (1): 101-106
87. Prazeres DM, Ferreira GN, Monteiro GA, et al. Large-scale production of pharmaceutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Trends Biotechnol, 1999, 17 (4): 169-174
88. Alex Eon-Duval, Karlene Gumbs, Christopher Ellett. Precipitation of RNA Impurities with High Salt in a Plasmid DNA Purification Process: Use of Experimental Design to Determine Reaction Conditions. Biotechnology and Bioengieering, 2003, 83 (5): 545-553
89. M.A.Teeters, S.E.Conrardy, B.L.Thomas, et al. Adsorptive membrane chromatography for purification of plasmid DNA. Journal of Chromatography A, 2003, 989 (1): 165-173
90. Per-Erik Gustavsson, Raf Lemmens, Tomas Nyhammar, et al. Purification of plasmid DNA with a new type of anion-exchange beads having a non-charged surface. Journal of Chromatography A, 2004, 1038 (1-2): 131-140
91. Rona Limor, Susan Gilad, Eina Kutikof, et al. Contaminant eluted from solid-phase plasmid affinity-purification protocol columns is not found using liquid-phase methods and can be prevented. Journal of biochemical and biophysical methods. 1999, 40 (1-2): 57-64
92. Dagmar Petsch and Friedrich Birger Anspach. Endotoxin removal from protein solutions. Journal of Biotechnology. 2000, 76 (2-3): 97-119
93. Judith Hellman, H. Shaw Warren. Antiendotoxin strategies. Infectious Disease Clinics of North America, 1999, 13 (2): 371-386
94. Liu S G, Tobias R, Mcclure, et al. Removal of endotoxin from recombinant proteins. Journal of Clinical Biochemistry, 1997, 30 (6): 455-160
95. Li Liping, Luo RG.. Quantitative determination of Ca2+ effects on endotoxin removal and protein yield in a two-stage ultrafilration process. Sep Sci Technol, 1999, 34 (9): 1729-1739
96. Anspach FB, Hilbeck O. Removal of endotoxin by affinity sorbents. Journal of chromatography A, 1995, 711 (1): 81-92
97. Minbo S, Watanabe T, Sato T, et al. Preparation of adsorbents for pyrogen adsorption. Journal of Chromatography, 1983, 248: 193-203
98. Phillip M. Montbriand, Robert W. Malone. Improved method for the removal of endotoxin from DNA. Journal of Biotechnology, 1996, 44 (1-3): 43-46
99. Mitzner S, Schneidewind J, Falkenhagen D, et al. Extracorporeal endotoxin removal by immobilized polyethylenimine. Artifical Organs, 1993, 17 (9): 775-781
100. Hirayama. C, Sakata. M, Nakamura. M, et al. Preparation of poly (Epsilon-lysine) adsorbents and application to selective removal of lipopolysaccharide. Journal of chromatography. B, 1999, 721: 187-195
101. Hirayama. C, Sakata. M, Ihara. H, et al. Effect of the pore size of an aminated poly (3-methyl L-glutamate) adsorbent on the selective removal of endotoxin. Anal Sci, 1992, 8: 805-510
102. M.S.Levy, I.J. Collins, J.T. Tsai, et al. Removal of contaminant nucleic acids by nitrocelluse filtration during pharmaceutical-grade plasmid DNA processing. Journal of Biotechnology, 2000, 76 (2-3): 197-205
103. Michael A. Winters, Jesse D. Richter, Sangeetha L.Sagar, et al. Plasmid DNA Purification by Selective Calcium Silicate Adsorption of Closely Related Impurities. Biotechnology Progress, 2003, 19 (2): 440-447
104. Prazeres, D., Jesse D., Collins, J.T., et al. Preparation purification of supercoiled plasmid DNA using anion-exchange chromatography. J.Chromatogr, 1998, 806: 31-45
105. Stahlberg.Jan. Retention models for ions in chromatography. Journal of Chromatography A, 1999, 855 (1): 3-12
106. 张思详, 韩娟. 凝胶色谱用于 DNA 检测的实验研究. 光谱仪器与分析, 2002, 2: 39-43
107. M. M. Diogo, J.A.Queiroz, G..A. Monterio, et al. Purification of a Cystic Fibrosis Plasmid Vector for Gene Therapy Using Hydrophobic Interaction Chromatography. Biotechnology and Bioengineering, 2000, 68 (5): 576-583
108. Yuan Li, Xiao-Yan Dong, Yan Sun. High-speed chromatographic purification of plasmid DNA with a customized biporous hydrophobic adsorbent. Biochemical Engineering Journal, 2005, 27 (1): 33-39
109. P Wils, V Escrious, A Warnery, et al. Efficient purification of plasmid DNA for gene transfer using triple-helix affinity chromatography. Gene Therapy, 1997, 4 (4): 323-330
110. John Woodgate, David Palfrey, David A. Nagel, et al. Protein-Mediated isolation of Plamsid DNA by a zinc Finger-Glutathione S-Transferase Affinity Linker. Biotechnology and Bioengineering, 2001, 79 (4): 450-456
111. Jan Kostal, Ashok Mulchandani, Wilfred Chen. Affinity Purification of Plasmid DNA by Temperature-Triggered Precipitation. Biotechnology and Bioengineering. 2004, 85 (3): 450-456
112. Lena M. Sandberg, Asa Bjurling, Philippe Busson, et al. Thiophilic interaction chromatography for supercoiled plasmid DNA purification. Journal of Biotechnology, 2004, 109 (1-2): 193-199
113. E.H. Chowdhury and Toshihiro Akaike. Rapid isolation of high quality, mulimeric plasmid DNA using zwitterionic detergent. Journal of Biotechnology, 2005, 119 (4): 343-347
114. Jason C., Murphy. Jamie A., Wibbenmeyer. George E. Fox, et al. Purification of plasmid DNA using selective precipitation by compaction agent. Nature Biotechnology, 1999, 17: 822-823
115. Hoopes, B.C and McClure. Studies on the selectivity of DNA precipitation by spermine. Nucl. Acids Res, 1981, 9 (20): 5493-5504
116. Cheng-Li Chiang, Ching-Shan Sung, Ting-Feng Wu, et al. Application of superparamagnetic nanoparticles in purification of plasmid DNA from bacterial cells. Journal of Chromatography B. 2005, 822 (1-2): 54-60
117. Cheng-Li Chiang and Ching-Shan Sung. Purification of transfection-grade plasmid DNA from bacterial cells with superparamagnetic nanoparticles. Journal of Magenetism and Magnetic Materials, 2005, 25: 189-199
118. Sindhu Balan, Jason Murphy, Igor Galaev, et al. Metal chelate affinity pricipitation of RNA and purification of plasmid DNA. Biotechnology Letters, 2003, 25: 1111-1116
119. Albertsson P-?. Partition of double-stranded and single-stranded deoxyribonucleic acid. Arch Biochem Biophys Supplement, 1962, 1: 264-270
120. Ohlsson R, Hentschel CC, Williams JG. A rapid method for the isolation of circular DNA using an aqueous two-phase partition system. Nucleic Acids Res, 1978, 5 (2): 583-590
121. Cole K D. Purification of plasmid and high molecular mass DNA using PEG-salt two-phase extraction. BioTechniques, 1991, 11 (1): 18, 20, 22-24
122. S.C.Ribeiro, G.A.Monterio., J.M.S.Cabral, et al. Isolation of Plasmid DNA from Cell Lysates by Aqueous Two-Phase Systems. Biotechnology and Bioengineering, 2002, 78 (4): 377-384
123. Inês P. Trindade, Maria M. Diogo, Duarte M. F. Prazeres, et al. Purification of plasmid DNAvectors by aqueous two-phase extraction and hydrophobic interaction chromatography. Journal of Chromatography A, 2003, 1082 (2): 176-184
124. Cecilia Kepka, Jenny Rhodin, Raf Lemmens, et al. Extraction of plasmid DNA from Escherichia coli cell lysate in a thermoseparating aqueous two-phase system. Journal of Chromatography A, 2004, 1024 (1-2): 95-104
125. M.A. Teeters, S.E. Conrardy, B.L. Thomas, et al. Adsorptive membrane chromatography for purification of plasmid DNA. Journal of Chromatography A, 2003, 989 (1): 165-173
126. Jochen Urthaler, Robert Schlegl, Ales Podgornik, et al. Application of monoliths for plasmid DNA purification Development and transfer to production. Journal of Chromatography A, 2005, 1065 (1): 93-106
127. Woff J.A., Malone R.W., Williams P., et al. Direct gene transfer into mouse muscle in vivo. Science, 1990, 247: 1465-1468
128. Mulligan RC. The basic science of gene therapy. Science, 1993, 260 (5110): 926-932
129. Guilherme N.M, Ferreira Gabriel A, Monterio, et al. Downstream processing of plasmid DNA for gene therapy and DNA vaccine applications. Tibech, 2001,18 (9): 380-388
130. Mor G., Klinman D.M., Shapiro S., et al. Complexity for the cytokine and antibody response elicited by immunizing mice with Plasmodium yoelii circumsporozoite protein plasmid DNA. J. Immunol, 1995, 155 (4): 2039-46
131. Moore AC, Hill. Progress in DNA-based heterologous prime-boost immunization strategies for malaria. Immunological Reviews, 2004, 199: 126-43
132. Sechi L A, Mara L, Cappai P, et al. Immunization with DNA vaccines encoding different mycobacterial antigens elicits a Th1 type immune response in lambs and protects against Mycobacterium avium subspecies paratuberculosis infection. Vaccine, 2006, 24 (3): 229-235
133. Martin T., Parker S.E., Hedstrom R., et al. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Human Gene Therapy, 1999, 10 (5): 759-768
134. Mac Gregor R. R., Boyer J. D., Ciccarelli R.B., et al. Safety and immune responses to a DNA-based human immunodeficiency virus (HIV) type I env/rev vaccine in HIV-infectedrecipients: follow-up data. J. Infect Dis., 2000, 181 (1): 406-412
135. Kanellos T., Sylvester I.D., Ambali A.G. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Human Gene Therapy, 1999, 10 (5): 759-768
136. Martin T., Parker S.E., Hedstrom R., et al. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Human gene therapy, 1999, 10 (5): 759-768
1. Kowalczyk D.W and Ertl H.C.J. Immune responses to DNA vaccines. Cell and Molecular Life Sciences, 1999, 55 (5):751-770
2. Sahota S S, Townsend M and Stevenson F K. Identification and assembly of V genes as idiotype-specific DNA fusion vaccines in multiple myeloma. Methods Mol Med, 2005, 113: 105-19
3. Rosati M, Von Gegerfelt A, Roth P, et al. DNA vaccines expressing different forms of simian immunodeficiency virus antigens decrease viremia upon SIVmac251 challenge. J Virol, 2005, 79 (13): 8480-92
4. Prazeres D.M.F. Large-scale production of pharmaceutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Trends in Biotechnology, 1999, 17 (4): 169-174
5. O’Kennedy R.D, Baldwin C and Keshavarz-Moore E. Effects of growth medium selection on plasmid DNA production and initial processing steps. Journal of Biotechnology, 2000, 76 (8):175-183
6. J.萨姆布鲁克, D.W 拉塞尔. 分子克隆实验指南第三版. 科学出版社
7. Wegrzyn G. Replication of plasmids during bacterial response to amino acid starvation. Plasmid, 1999, 41(1): 1-16
8. Kristala J. P, Sangeetha S, Jason M, et.al. Industrial scale production of plasmid DNA for vaccine and gene therapy: plasmid design, production, and purification. Enzyme and Microbial Technology, 2003, 16 (33): 865–883
1. W Chen. Automated fed-batch fermentation with feed-back controls based on dissolved oxygen (DO) and pH for prouuction of DNA vaccines. Journal of Industrial Microbiology & Biotechnology, 1997, 18 (1): 43-48
2. J.萨姆布鲁克, D.W. 拉塞尔. 分子克隆第三版
3. R.O’Kennedy C.J, Houghton. Effects of growth environment on recombinant plasmid stabilityin Saccharomyces cersvisiae grown in continuous culture. Appl Microbiol Biotechnol, 1995, 44 (1-2): 126-132
4. Corchero J L, Villaverde A. Plasmid maintenance in Escherichia coli recombinant cultures is dra, steadily, and specifically influenced by features of the encoded proteins. Biotechnology and Bioengineering, 1998, 58 (6): 625-632
5. Lahijani R, Hulley G. High-yield production of PBR322-derived plasmids intended for human gene therapy by employing a temperature controllable point mutation. Human Gene Therapy, 1996, 7 (16): 1971-80
6. D.J.Korz. Simple fed-batch technique for high cell density cultivation of Escherichia coli. Journal of Biotechnology, 1995, 39 (1): 59-65
7. Reinikainen P, Korpela K. Escherichia coli production in fermentor. Biotechnology and Bioengineering, 1989, 33: 386–93
8. Kristala Jones Prather. Industrial scale production of plasmid DNA for vaccine and gene therapy: plasmid design, production, and purification. Enzyme and Microbial Technology, 2003, 33 (7): 865-883
1. 荣晓花, 凌沛学. 溶菌酶的研究进展. 中国生化药物杂志, 1999, 20 (6): 916-919
2. 史萸芳. 溶菌酶抑菌作用的实验. 牙体牙髓牙周病学杂志, 1996, 6 (1): 26-28
3. 防治奶牛乳腺炎的基因药物. 专利号:ZL02148545.3
4. Sambrook J, Russell DW. Molecular Cloning A Laboratory Manual. Third Edition, New York: Cold Spring Harbor Laboratory Press, 2002
5. 柳 湘, 田培坤, 顾键人, 等. 非病毒载体系统在基因治疗中的应用. 国外医学肿瘤学分册, 2001, 28 (3): 189-191
6. Kristala Jones Prather, Sangeetha Sagar, Jason Murphy, et al. Industrial scale production of plasmid DNA for vaccine and gene therapy: plasmid design, production, and purification. Enzyme and Microbial Technology, 2003, 33 (7): 865-883
7. Ferber D. Gene therapy: safer and virus free. Science, 2001, 294 (5547): 1638-42
8. Guilherme N.M. Ferreira, Gabriel A. Monteiro, Duarte M.F, et al. Downstream processing of plasmid DNA for gene therapy and DNA vaccine applications. Trends Biotechnol, 2000, 18 (9): 380-388
9. J.Zhang R. Greasham. Chemically defined media for commercial fermentation. Appl Microbiol Biotechnol, 1999, 51: 407-421
10. 陈天寿. 微生物培养基的制造与应用. 中国农业出版社, 1995
1. 薛方明. 防治奶牛乳腺炎基因药物的临床试验与安全平均. 扬州大学学位论文, 2005
2. Birnboim, H.C. and Doly. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res, 1979, 7 (6): 1513-23
1. DuarteM.F.Prazeres, Guilherme N. M. Ferreira, GabrielA. Monteiro, et al. Large-scale production of pharmaeutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Tibtech Appril, 1999, 19 (2): 169-174
2. Duarte Miguel F. Prazeres, Thomas Schluep, Charles Cooney, et al. Preparative purification of supercoiled plasmid DNA using anionexchange chromatography. Journal of Chromatography A, 1998, 806 (1): 31- 45
3. Tseng WC, Ho FL. Enhanced purification of plasmid DNA using Q-Sepharose by modulationof alcohol concentrations. J Chromatogr B Analyt Technol Biomed Life Sci, 2003, 791(1-2): 263-72
4. Eon-Duval A, Burke G. Purification of pharmaceutical-grade plasmid DNA by anion-exchange chromatography in an RNase-free process. J Chromatogr B Analyt Technol Biomed Life Sci, 2004, 804 (2): 327-35
5. Darby RA, Hine AV. LacI-mediated sequence-specific affinity purification of plasmid DNA for therapeutic applications. FASEB J, 2005, 19 (7): 801-3
6. Wahlund PO, Gustavsson PE, Izumrudov VA, et al. Precipitation by polycation as capture step in purification of plasmid DNA from a clarified lysate. Biotechnol Bioeng, 2004, 87 (5): 675-84
7. Shenoy SR, Khalid M, Gupta A, et al. Purification of an Enterobacter aerogenes plasmid DNA using MnCl2 as compaction agent. Anal Biochem, 2003, 321 (2): 256-8
8. Balan S, Murphy J, Galaev I, et al. Metal chelate affinity precipitation of RNA and purification of plasmid DNA. Biotechnol Lett, 2003, 25 (13): 1111-6
9. J.萨姆布鲁克, D.W 拉塞尔, 分子克隆实验指南三版, 科学出版社, 2003
10. M. Susana Levy, Ronan D. O’Kennedy, Parviz Ayazi-Shamlou, et al. Biochemical engineering approaches to the challenges of producing pure plasmid DNA. Trends Biotechnol, 2000, 18 (7): 296-305
11. Alex Eon-Duval, RobertH.MacDuff, Carol A.Fisher, et al. Removal of RNA impurities by tangential flow filtration in an RNase-free plasmid DNA purification process. Analytical Biochemistry, 2003, 316 (2): 66-73
12. Eon-Duval, Gumbs K, Ellett C. Precipitation of RNA impurities with high salt in a plasmid DNA purification process: use of experimental design to determine reaction conditions. Biotechnol Bioeng, 2003, 83 (5): 544-53
13. M.S. Levy, I.J. Collins, J.T. Tsai P., et al. Removal of contaminant nucleic acids by nitrocellulose filtration during pharmaceutical-grade plasmid DNA processing. Journal of Biotechnology, 2000, 76 (2-3): 197-205
14. Lander RJ, Winters MA, Meacle FJ, et al. Fractional precipitation of plasmid DNA from lysateby CTAB. Biotechnol Bioeng, 2002, 79 (7): 776-84
15. Teeters MA, Conrardy SE, Thomas BL. Adsorptive membrane chromatography for purification of plasmid DNA. J Chromatogr A, 2003, 989 (1): 165-73
16. Anspach FB. Endotoxin removal by affinity sorbents. J Biochem Biophys Methods, 2001, 49 (1-3): 665-81
1. D. W. Kowalczyk, H. C. Ertl. Immune response to DNA vaccines. Cellular and Molecular Life Science, 1999, 55: 751-770
2. Robertson J, Griffiths E. WHO Guidelines for Assuring the Quality of DNA Vaccines, Biologicals, 1998, 26 (3): 205-212
3. Guilherme N.M.Ferrira, Gabriel A. Monteiro, Duarte M.F, et al. Downstream processing of plasmid DNA for gene therapy and DNA vaccine applications. Trends Biotechnol, 2000, 18 (9): 380-388
4. May CM, Laurent C, Jean M. A novel potent strategy for gene delivery using a single peptide vector as a carrier. Nucleic Acids Research, 1999, 27 (17): 3510-3517
5. Schuster M, Wu GY, Walton CM. Multicoponent DNA carrier with a vesicular stomatitis virus G-peptide greatly enhances liver targeted gene expression in mice. Bioconjug Chem, 1999, 10 (6): 1075-1083
6. QIAGEN Plasmid Purification Handbook, 2003: 72
7. Martich GD, Boujoukos AJ, Suffredini AF. Response of man to endotoxin. Immunobiology, 1993, 187 (3-5): 403-16
8. Bragonzi A, Boletta A, Motexs A, et al. Comparison between cationic polymers and lipids in mediating systemic gene delivery to the drugs. Gene Ther, 1999, 6 (12): 1995-2004.
9. 王风阳, 孟庆文, 扈容良, 等. 含 LacZ 重组逆转录病毒的制备及其在 NIH3T3 细胞的表达. 中国预防兽医学报, 2001, 23 (5): 329-331
10. 钱锋, 肖成祖. 影响非洲绿猴肾细胞脂质体转染效率的因素. 生物技术通报, 1998, 5: 31-35
11. 廖文俊, 刘彦仿, 刘玉峰. β-半乳糖苷酶基因转染角质形成细胞的瞬时表达检测. 临床皮肤科杂志, 1999, 28 (2): 71-74
12. Kichler A,Leborgne C, Coeytauxe, et al. Polyethylenimine-mediated gene delivery: a mechanistic study. J Gene Med, 2001, 3 (2): 135-144
13. Koping-Hoggard M, Tubuleka I, Guan H, et al. Chitosan as a nonviral gene delivery system structure-property relationships and characteristics compared with polyethylenimine in vitro and after lung administration in vivo. Gene Ther, 2001, 8 (14): 1108-1112
14. Zaumer W, Brunner S, Buschle M, et al. Differential behavior of lipid based and polycation based gene transfer systems in transfecting primary human fibroblasts a potentianl role of polylysine in nuclear transport. Biochim Biophys Acta, 1999, 1428 (1): 57-61
15. Durocher Y, Perret S, Kamen A. High-level and high-throughout recombinant protein production by transient transfection of suspension growing human 293-EBNA1 cells. Nucleic Acids Research, 2002, 30: 1-9
16. 莫国玉, 陈光明, 黄芝瑛, 等. 治疗型 HBV DNA 疫苗在小鼠组织中的分布和长期毒性研究. 解放军医学杂志, 2003, 28 (6): 504-507
17. 李经忠, 王青青. 非病毒载体的研究现状. 国外医学分子生物学分册, 2002, 24 (5): 317-320
18. Ferreira. Plasmid manufacturing-an overview. In Plasmids for Therapy and Vaccination, Schleef M (ed). Whiley-VCH: Weinheim, 2001, 203-204
19. Raetz CR. Biochemistry of endotoxins. Annu Rev Biochem, 1990, 59: 129-170
20. Kastowsky M, Gutberler T, Bradaczek H. Molecular modeling of the three-dimensional structure and conformational flexibility of bacterial lipopolysaccharide. J Bacteriol, 1992,174: 4798-806
1. 莫国玉, 陈光明, 黄芝瑛, 等. 治疗型 HBV DNA 疫苗在小鼠组织中的分布和长期毒性研究. 解放军医学杂志, 2003, 28 (6): 504-507
2. John Donnelly, Karin Berry, Jeffrey B. Ulmer. Technical and regulatory hurdles for DNA vaccines. International Journal for Parasitology, 2003, 33 (5-6): 457-467
3. Kowakczyk DW and Ertl HC. Immune responses to DNA vaccines. Cellular and Molecular Life Sciences, 1999, 55 (5): 751-770
4. Tang D. C., Devit M. and Johnston S. A. Genetic immunization is a simple method for eliciting an immune response. Nature, 1992, 356 (6365): 152-154
5. Prazeres D.M.F. Large-scale production of pharmaceutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Trends in Biotechnology, 1999, 17 (4): 169-174
6. QIAGEN Plasmid Purification Handbook, 2003
7. Robertson J, Griffiths E. WHO Guidelines for Assuring the Quality of DNA Vaccines. Biologicals, 1998, 26 (3): 205-212
8. Dagmar Petsch, Friedrich Birger Anspach. Endotoxin removal from protein solutions. Journal of Biotechnology, 2000, 76 (2-3): 97-119
9. Lydia A. Falk, Leslie K. Ball. Current status and future trends in vaccine regulation-USA. Vaccine, 2001, 19 (13-14): 1567-1572
10. J.萨姆布鲁克, D.W 拉塞尔, 分子克隆实验指南第三版, 科学出版社
1. Verfailliea T, Cox B.M. Goddeeris Immunostimulatory capacity of DNA vaccine vectors in porcine PBMC: a specific role for CpG-motifs. Veterinary Immunology and Immunopathology, 2005, 103 (1-2): 141-151
2. Klinman D M. CpG DNA as a vaccine adjuvant. Expert Rev Vaccines, 2003, 2 (2): 305-15
3. 莫国玉, 陈光明, 黄芝瑛, 等. 治疗型HBV DNA疫苗在小鼠组织中的分布和长期毒性研究. 解放军医学杂志, 2003, 28 (6): 504-507
4. Guidance for Industry Considerations for Plasmid DNA Vaccines for Infectious Disease Indications
5. Wang Z, Troilo P J, Wang X, et al. Detection of integration of plasmid DNA into host genomic DNA following intramuscular injection and electroporation. Gene Therapy, 2004, 11 (8): 711-21
6. Ledwith B J, Manam S, Troilo P J, et al. Plasmid DNA vaccines: assay for integration into host genomic DNA. Dev Biol (Basel), 2000, 104: 33-43
7. Liu Y, Liggitt D, Zhong W, et al. Cationic liposome-mediated intravenous gene delivery. Biol Chem, 1995, 270 (42): 24864-70
8. Mahato K, Kawabata Y, Takakura M, et al. In vivo disposition characteristics of plasmid DNA complexed with cationic liposomes. J Drug Target, 1995, 3 (2): 149-157
9. McLean J.W, Fox E A, Baluk P, et al. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am J Physiol, 1997, 273: H387-404
10. Delphine Lechardeura A.S, Verkmanb and Gergely L. Lukacs. Intracellular routing of plasmid DNA during non-viral gene transfer. Advanced Drug Delivery Reviews, 2005, 57 (5): 755-767
11. Nichols WW, Ledwith BJ, Manam SV, et al. Potential DNA vaccine integration into host cell genome. Ann Nei York Acad Sci, 1995, 772: 30-39
2. 吴乃虎. 基因工程原理. 科学出版社(第二版), 2001
3. Carole L. Bassett and Sidney R. Kushner. Exonucelases I, III and V are required for stability of ColE1-Related Plasmids in Escherichia coli. Journarl of Bacteriology, 1984, 157 (2): 661-664
4. Joachim Stadler, Raf Lemmens and Tomas Nyhammar. Plasmid DNA purification. The Journal of Gene Medicine, 2004, 6 (1): S54-66
5. 张玉静 主编. 分子遗传学. 科学出版社, 2002
6. J.萨姆布鲁克, D.W 拉塞尔. 分子克隆实验指南第三版. 科学出版社, 2003
7. Wang Z, Yuan Z, Hengge UR. Processing of plasmid DNA with ColE1-like replication origin. Plasmid, 2004, 51 (3): 149-61
8. Chambers S, Barstow D, Minton N. The pMTL nic-cloning vectors I Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene, 1988, 68 (1): 139-49
9. Yavachev L, Ivanov I. What does the homology between E. coli tRNAs and RNAs controlling ColE1 plasmid replication means?. J. Theor. Biol, 1988, 131 (2): 235-241
10. Wang,Z, Le,G, Shi Y, et al. A model for regulation of ColE1-like plasmid replication by uncharged tRNAs in amino acid-starved Escherichia coli cells. Plasmid, 2002c, 47 (2): 69-78
11. Minton NP. Improved plasmid vectors for the isolation of translational lac gene fusions. Gene, 1984, 31 (1-3): 269-73
12. N.A. Horn, J.A. Meek, G. Budahazi, M. Cancer gene therapy using plasmid DNA: purificationof DNA for human clinical trials. Human Gene Therapy, 1995, 6 (5): 565-573
13. N.C. Wan, J.C. Goodrick. Method for increasing plasmid yield. United States Patent#5, 1996, 487, 986
14. R. Lahijani, G. Hulley, G. Soriano, N.A. Horn, et al. High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature-controllable point mutation. Hum Gene Ther, 1996, 7 (16): 1971-80
15. Uhlin B, Nordstr?m K. A runaway-replication mutant of plasmid R1drd-19: temperature-dependent loss of copy number control. Mol Gen Genet, 1978, 165 (2): 167-79
16. Remaut E, Tsao H, Fiers W. Improved plasmid vectors with a thermo-inducible expression and temperature-regulated runaway replication. Gene, 1983, 22 (1): 103-13
17. Ansorge MB, Kula MR. Investigating expression systems for the stable large-scale production of recombinant leucine-dehydrogenase from Bacillus cereus in Escherichia coli. Appl Microbiol Biotechnol, 2000, 53 (6): 668-73
18. Chao Y, Chern J, Wen C. Coupling the T7 A1 promoter to the runaway-replication vector as an efficient method for stringent control and high-level expression of lacZ. Biotechnol Prog, 2001, 17 (1): 203-7
19. Kristala Jones Prather, Sangeetha Sagar, Jason Murphy, et al. Industrial scale production of plasmid DNA for vaccine and gene therapy: plamsid design, production, and purification. Enzyme and Microbial Technology, 2003, 33 (7): 865-883
20. Smith H. Regulatory considerations for nucleic acid vaccines. Vaccine, 1994, 12 (16): 1515-9
21. Gurunathan S, Klinman D, Seder R. DNA vaccines: immunology, application, and optimization. Annu Rev Immunol, 2000, 18: 927-74
22. Davis M, Huang E. Transfer and expression of plasmids containing human cytomegalovirus immediate-early gene promoter-enhancer sequences in eukaryotic and prokaryotic cells. Biotechnol Appl Biochem, 1988, 10 (1): 6-12
23. Chapman B, Thayer R, Vincent K, et al. Effect of intron A from human cytomegalovirus (Towne) immediate-early gene on heterologous expression in mammalian cells. Nucleic AcidsResearch, 1991, 19 (14): 3979-86
24. Verfailliea T, Cox B.M. Goddeeris. Immunostimulatory capacity of DNA vaccine vectors in porcine PBMC: a specific role for CpG-motifs. Veterinary Immunology and Immunopathology, 2005, 103 (1-2): 141-151
25. Klinman D M. CpG DNA as a vaccine adjuvant. Expert Rev Vaccines, 2003, 2 (2): 305-15
26. FDA. Points to consider on plasmid DNA vaccines for preventive infectious diseases. 1996, Docket No. 96N-0400
27. Robertson J, Griffiths E. WHO guidelines for assuring the quality of DNA vaccines. Biologicals, 1998, 26 (3): 205-12
28. Falk LA, Ball LK. Current status and future trends in vaccine regulation--USA. Vaccine, 2001, 19 (13-14): 1567-72
29. Cichutek K. DNA vaccines: development, standardization and regulation. Intervirology, 2000, 43 (4-6): 331-8
30. Robertson JS, Cichutek K. European union guidance on the quality, safety and efficacy of DNA vaccines and regulatory requirements. Dev Biol, 2000, 104: 53-6
31. Smith HA. Regulatory considerations for nucleic acid vaccines. Vaccine, 1994, 12 (16): 1515-9
32. Smith HA. Regulation and review of DNA vaccine products. Dev Biol, 2000, 104: 57-62
33. W Chen, C Graham and RB Ciccarelli. Automated fed-batch fermentation with feed-back controls based on dissolved oxygen (DO) and pH for production of DNA vaccines. Journal of industrial Microbiology and Biotechnology, 1997, 18 (1): 43-48
34. 赵 霞, 叶 勤, 俞俊棠. 影响重组工程菌发酵产率的因素. 工业微生物, 1995,25(2): 26-32
35. Erik Remaut, Hsu Tsao and Walter Fiers. Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication. Gene, 1983, 22 (1): 103-113
36. Robin-Ann Klotsky and Ira Schwartz. Measurement of cat expression from growth-rate-regulated promoters employing β-lactamase activity as an indicator of plasmid copy number. Gene, 1989, 55 (1): 141-146
37. 邱英华, 沈 益, 王玉海, 等. Cecropin X 发酵过程中工程菌质粒稳定性的研究. 微生物学报, 2004, 44 (3): 390-391
38. Jos ML Secgers, Cristian M Franke, Rense K, et al. Use of continuous culture for the selection of plasmids with improved segregational stability. Plasmid, 1995, 33 (1): 71-77
39. Nordstrom K, Molin S, Aagaard-Hansen H. Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid, 1980, 4 (2): 215-27
40. Engberg B and Nordstrom, K. J. Plasmid stability in E.coli. J. Bacteriol, 1975, 179: 123
41. Futchjer AB, Cox BS. Copy number and the stability of 2-micron circle-based artificial plasmids of Saccharomyces cerevisiae. J.Bacteriol, 1984, 157 (1): 283-90
42. Caulcott CA, Dunn A, Robertson HA, et al. Investigation of the effect of growth environment on the stability of low-copy-number plasmids in Escherichia coli. J Gen Microbiol, 1987, 133 (7): 1881-9
43. 杨海燕, 方呈祥, 张珞珍, 等. 不同质粒在大肠杆菌宿主细胞中的不稳定性研究. 河南农业大学学报, 1999, 33 (2): 190-193
44. Chang LL, Hwang LY and Hwang CF, et al. Study of high density Escherichia coli fermentation for production of porcine somatotropin protein. Ann N Y Acad Sci, 1991, 27 (5): 259-72
45. Ross H. Durland, Eric M. Eastman. Manufacturing and quality control of plasmid-based gene expression systems. Advanced Drug Delivery Reviews, 1998, 2 (30): 33-41
46. Takapa 商品目录, 2004-2005
47. 许乃用. 稳定质粒拷贝数提高重组蛋白质的产量. 工业微生物, 2002, 32 (2): 58
48. 姜泊, 张亚力. 分子生物学常用试验方法. 北京: 人民军医出版社, 1996
49. 卢圣栋. 现代分子生物学实验技术. 北京: 高等教育出版社, 1999, 54
50. Lahijani R, Hulley G, Soriano G, et al. High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature controllable point mutation. Human Gene Therapy, 1996, 7 (16): 1971-80
51. Schmidt T, Friehs K, Schleef M, et al. In-process analysis of plasmid copy number for fermentation control. Pacesetter, 2001, 5 (1): 4-6
52. Korz DJ, Rinas U, Hellmuth K, et al. Simple fed-batch technique for high cell densitycultivation of Escherichia coli. J Biotechnol, 1995, 39 (1): 59-65
53. O’Kennedy RD, Ward JM, Keshavarz-Moore E. Effects of fermentation strategy on the characteristics of plasmid DNA production. Biotechnol Appl Biochem, 2003, 37 (1): 83-90
54. Ronan D. O’Kennedy, John M. Ward and Eli Keshavarz-Moore. Effects of fermentation stategy on the characteristics of plasmid DNA production. Biotechol. Appl. Biochem, 2003, 37: 83-90
55. R.D. O’Kennedy, C.Baldwin, E.Keshavarz-Moore. Effects of growth medium selection on plasmid DNA production and initial processing steps. Journal of Biotechnology, 2000, 76 (2-3): 175-183
56. 俞俊棠, 唐孝宣. 生物工艺学. 上海: 华东理工大学出版社, 2000: 211
57. Nakasu S and Tomizawa J. Structure of the ColE1 DNA molecule before segregation to daughter molecules. Proc Natl Acad Sci U S A, 1992, 89 (21): 10139-43
58. D.J. Rorz, U. Rinas, K. Hellmuth, et al. Simple fed-batch technique for high density cultivation of Escherichia coli. Journal of Biotechnology, 1995, 39 (1): 59-65
59. Lahijani R, Hulley G, Soriano G, et al. High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature-controllable point mutation. Hum Gene Ther, 1996, 7 (16): 1971-80
60. Reinikainen P, K Korpela, V Nissine, et al. Escherichia coli plasmid production in fermentator. Biotechnol Bioeng, 1989, 33: 386-393
61. QIAGEN Plasmid Purification Handbook, Second Edition, August 2003
62. Michel Thiry and Doriano Cingotanl. Optimizing scale-up fermentation processes. Trends in Biotechnology, 2002, 20 (3): 103-105
63. Lee SY. High cell-density culture of Escherichia coli Reviews. Trends Biotechnol, 1996, 14 (3): 98-105
64. Docket NO.96N-0400 Points to Consider on Plasmid DNA Vaccines for Preventive Infectious Disease Indications
65. Ellenberg SS. Safety considerations for new vaccine development. Pharmacoepidemiol Drug Saf, 2001, 10 (5): 411-5
66. D.M.F. Prazeres, G.N.M.Ferreira. Design of flowsheets for the recovery and purification of plasmids for gene therapy and DNA vaccination. Chemical Engineering and Processing, 2003 , 4 (43): 615-630
67. D.Petrides. Bioprocess design in: R.G. Harrison. P.W. Todd, S.R.Rudge, D. Petrides (eds). Biseparations Scinece and Engineering, Oxford University Press,Oxford, UK, 2003, pp. 319-372
68. 郭利平, 章佩芬, 唐青涛. 基因治疗和 DNA 疫苗质粒 DNA 的下游加工技术. 华西药学杂志, 2002, 17 (2): 159-162
69. Zhijun Wang, Guowei Le, Yonghui Shi, et al. Studies on recovery plasmid DNA from Echerichia coli by heat treatment. Process Biochemistry, 2002, 38: 199-206
70. Birnboim HC, Doly J. rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res, 1979, 7 (6): 1513-23
71. Jon R. Sayers, Debbie Evans and James B. Thomson. Identification and Eradication of a Denatured DNA Isolated during Alkaline Lysis-Based Plasmid Purification Procedures. Analytical Biochemistry, 1996, 241 (2): 186-189
72. Jun Song, Christian Geltinger and Kailai Sun, et al. Direct Lysis Method for the Rapaid Preparation of Plasmid DNA. Analytical Biochemistry, 1999, 271 (1): 89-91
73. Wen-Chi Tseng, Fu-lyn Ho, Tsuei-Yun Fang, et al. Effect of alcohol on purification of plasmid DNA using ion-exchange membrane. Journal of Membrane Scinece, 2004, 233: 161-167
74. Ferreira GNM. A comparison of gel filtration chromatographic supports for plasmid purification. Biotechnol Techn, 1997, 11: 417-420
75. Diogo MM, Queiroz JA, Monteiro GA, et al. Separation and analysis of denatured plasmid forms using hydrophobic interaction chromatography. Anal Biochem, 1999, 275 (1): 122-124
76. Matteo D. Costioli, lgor Fisch, Frank Hilbrig, et al. DNA purification by Triple-Helix Affinity Precipitation. Biotechnology and Bioengineering, 2003, 81 (5): 535-545
77. Alex Eon-Duval, Rober H. MacDuff, Carol A, et al. Removal of RNA impurities by tangential flow filtration in an RNase-free plasmid DNA purification process. Analytical Biochemistry,2003, 316 (1): 66-73
78. Dan V. Mourich, Michael W. Munks, Jason C. Murphy, et al. Spermine compaction is an efficient and ecomomical method of producing vaccination-grade DNA. Journal of Immunological Methods, 2003, 274 (1-2): 257-264
79. Russel J.Lander, Michael A. Winters, Francis J. Meacle, et al. Fractional Precipitation of Plasmid DNA from Lysate by CTAB. Biotechnology and Bioengineering, 2002, 79 (7): 777-784
80. Per-Olof Wahlund, Per-Erik Gustavsson, Vladimir A. Izumrudov, et al. Polyelectrolyte complexes as a tool for purification of plasmid DNA Background and development. Journal of Chromatograohy B, 2004 , 807 (1): 121-127
81. Raf Lemmens, Urban Olsson, Tomas Nyhammar, et al. Supercoiled plasmid DNA: selective purification by thiophilic/aromatic adsorption. Journal of Chromatography B, 2003, 784 (2): 291-300
82. Sandhya R. Shenoy, Mohammad Khalid, Anshu Gupta, et al. Purification of an Enterobacter aerogenes Plasmid DNA using MnCl2 as compaction agent. Analytical Biochemistry, 2003, 321 (2): 256-258
83. B. Atkinson, F. Mavituna (Eds). Biochemical Engineering and Biotechnology Handbook, Macmillan, New York, USA, 1991
84. Sofer, G, and Hagel, L. Handbook of Process Chromatography: A Guide to Optimization, scale-up and Validation (1st edn), Academic Press
85. G.D.Cooke, R.M.Cranenburgh, J.A.J. Hanak, et al. Purification of essentially RNA free plasmid DNA using a modified Escherichia coli host strain expressing ribonuclease A. Journal of Biotechnology, 2001, 85 (3): 297-304
86. David W.Kahn, Michelle D. Butler, Darien L. Cohen, et al. Purification of Plasmid DNA by Tangential Flow Filtration. Biotechnology and Bioengineering, 2000, 69 (1): 101-106
87. Prazeres DM, Ferreira GN, Monteiro GA, et al. Large-scale production of pharmaceutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Trends Biotechnol, 1999, 17 (4): 169-174
88. Alex Eon-Duval, Karlene Gumbs, Christopher Ellett. Precipitation of RNA Impurities with High Salt in a Plasmid DNA Purification Process: Use of Experimental Design to Determine Reaction Conditions. Biotechnology and Bioengieering, 2003, 83 (5): 545-553
89. M.A.Teeters, S.E.Conrardy, B.L.Thomas, et al. Adsorptive membrane chromatography for purification of plasmid DNA. Journal of Chromatography A, 2003, 989 (1): 165-173
90. Per-Erik Gustavsson, Raf Lemmens, Tomas Nyhammar, et al. Purification of plasmid DNA with a new type of anion-exchange beads having a non-charged surface. Journal of Chromatography A, 2004, 1038 (1-2): 131-140
91. Rona Limor, Susan Gilad, Eina Kutikof, et al. Contaminant eluted from solid-phase plasmid affinity-purification protocol columns is not found using liquid-phase methods and can be prevented. Journal of biochemical and biophysical methods. 1999, 40 (1-2): 57-64
92. Dagmar Petsch and Friedrich Birger Anspach. Endotoxin removal from protein solutions. Journal of Biotechnology. 2000, 76 (2-3): 97-119
93. Judith Hellman, H. Shaw Warren. Antiendotoxin strategies. Infectious Disease Clinics of North America, 1999, 13 (2): 371-386
94. Liu S G, Tobias R, Mcclure, et al. Removal of endotoxin from recombinant proteins. Journal of Clinical Biochemistry, 1997, 30 (6): 455-160
95. Li Liping, Luo RG.. Quantitative determination of Ca2+ effects on endotoxin removal and protein yield in a two-stage ultrafilration process. Sep Sci Technol, 1999, 34 (9): 1729-1739
96. Anspach FB, Hilbeck O. Removal of endotoxin by affinity sorbents. Journal of chromatography A, 1995, 711 (1): 81-92
97. Minbo S, Watanabe T, Sato T, et al. Preparation of adsorbents for pyrogen adsorption. Journal of Chromatography, 1983, 248: 193-203
98. Phillip M. Montbriand, Robert W. Malone. Improved method for the removal of endotoxin from DNA. Journal of Biotechnology, 1996, 44 (1-3): 43-46
99. Mitzner S, Schneidewind J, Falkenhagen D, et al. Extracorporeal endotoxin removal by immobilized polyethylenimine. Artifical Organs, 1993, 17 (9): 775-781
100. Hirayama. C, Sakata. M, Nakamura. M, et al. Preparation of poly (Epsilon-lysine) adsorbents and application to selective removal of lipopolysaccharide. Journal of chromatography. B, 1999, 721: 187-195
101. Hirayama. C, Sakata. M, Ihara. H, et al. Effect of the pore size of an aminated poly (3-methyl L-glutamate) adsorbent on the selective removal of endotoxin. Anal Sci, 1992, 8: 805-510
102. M.S.Levy, I.J. Collins, J.T. Tsai, et al. Removal of contaminant nucleic acids by nitrocelluse filtration during pharmaceutical-grade plasmid DNA processing. Journal of Biotechnology, 2000, 76 (2-3): 197-205
103. Michael A. Winters, Jesse D. Richter, Sangeetha L.Sagar, et al. Plasmid DNA Purification by Selective Calcium Silicate Adsorption of Closely Related Impurities. Biotechnology Progress, 2003, 19 (2): 440-447
104. Prazeres, D., Jesse D., Collins, J.T., et al. Preparation purification of supercoiled plasmid DNA using anion-exchange chromatography. J.Chromatogr, 1998, 806: 31-45
105. Stahlberg.Jan. Retention models for ions in chromatography. Journal of Chromatography A, 1999, 855 (1): 3-12
106. 张思详, 韩娟. 凝胶色谱用于 DNA 检测的实验研究. 光谱仪器与分析, 2002, 2: 39-43
107. M. M. Diogo, J.A.Queiroz, G..A. Monterio, et al. Purification of a Cystic Fibrosis Plasmid Vector for Gene Therapy Using Hydrophobic Interaction Chromatography. Biotechnology and Bioengineering, 2000, 68 (5): 576-583
108. Yuan Li, Xiao-Yan Dong, Yan Sun. High-speed chromatographic purification of plasmid DNA with a customized biporous hydrophobic adsorbent. Biochemical Engineering Journal, 2005, 27 (1): 33-39
109. P Wils, V Escrious, A Warnery, et al. Efficient purification of plasmid DNA for gene transfer using triple-helix affinity chromatography. Gene Therapy, 1997, 4 (4): 323-330
110. John Woodgate, David Palfrey, David A. Nagel, et al. Protein-Mediated isolation of Plamsid DNA by a zinc Finger-Glutathione S-Transferase Affinity Linker. Biotechnology and Bioengineering, 2001, 79 (4): 450-456
111. Jan Kostal, Ashok Mulchandani, Wilfred Chen. Affinity Purification of Plasmid DNA by Temperature-Triggered Precipitation. Biotechnology and Bioengineering. 2004, 85 (3): 450-456
112. Lena M. Sandberg, Asa Bjurling, Philippe Busson, et al. Thiophilic interaction chromatography for supercoiled plasmid DNA purification. Journal of Biotechnology, 2004, 109 (1-2): 193-199
113. E.H. Chowdhury and Toshihiro Akaike. Rapid isolation of high quality, mulimeric plasmid DNA using zwitterionic detergent. Journal of Biotechnology, 2005, 119 (4): 343-347
114. Jason C., Murphy. Jamie A., Wibbenmeyer. George E. Fox, et al. Purification of plasmid DNA using selective precipitation by compaction agent. Nature Biotechnology, 1999, 17: 822-823
115. Hoopes, B.C and McClure. Studies on the selectivity of DNA precipitation by spermine. Nucl. Acids Res, 1981, 9 (20): 5493-5504
116. Cheng-Li Chiang, Ching-Shan Sung, Ting-Feng Wu, et al. Application of superparamagnetic nanoparticles in purification of plasmid DNA from bacterial cells. Journal of Chromatography B. 2005, 822 (1-2): 54-60
117. Cheng-Li Chiang and Ching-Shan Sung. Purification of transfection-grade plasmid DNA from bacterial cells with superparamagnetic nanoparticles. Journal of Magenetism and Magnetic Materials, 2005, 25: 189-199
118. Sindhu Balan, Jason Murphy, Igor Galaev, et al. Metal chelate affinity pricipitation of RNA and purification of plasmid DNA. Biotechnology Letters, 2003, 25: 1111-1116
119. Albertsson P-?. Partition of double-stranded and single-stranded deoxyribonucleic acid. Arch Biochem Biophys Supplement, 1962, 1: 264-270
120. Ohlsson R, Hentschel CC, Williams JG. A rapid method for the isolation of circular DNA using an aqueous two-phase partition system. Nucleic Acids Res, 1978, 5 (2): 583-590
121. Cole K D. Purification of plasmid and high molecular mass DNA using PEG-salt two-phase extraction. BioTechniques, 1991, 11 (1): 18, 20, 22-24
122. S.C.Ribeiro, G.A.Monterio., J.M.S.Cabral, et al. Isolation of Plasmid DNA from Cell Lysates by Aqueous Two-Phase Systems. Biotechnology and Bioengineering, 2002, 78 (4): 377-384
123. Inês P. Trindade, Maria M. Diogo, Duarte M. F. Prazeres, et al. Purification of plasmid DNAvectors by aqueous two-phase extraction and hydrophobic interaction chromatography. Journal of Chromatography A, 2003, 1082 (2): 176-184
124. Cecilia Kepka, Jenny Rhodin, Raf Lemmens, et al. Extraction of plasmid DNA from Escherichia coli cell lysate in a thermoseparating aqueous two-phase system. Journal of Chromatography A, 2004, 1024 (1-2): 95-104
125. M.A. Teeters, S.E. Conrardy, B.L. Thomas, et al. Adsorptive membrane chromatography for purification of plasmid DNA. Journal of Chromatography A, 2003, 989 (1): 165-173
126. Jochen Urthaler, Robert Schlegl, Ales Podgornik, et al. Application of monoliths for plasmid DNA purification Development and transfer to production. Journal of Chromatography A, 2005, 1065 (1): 93-106
127. Woff J.A., Malone R.W., Williams P., et al. Direct gene transfer into mouse muscle in vivo. Science, 1990, 247: 1465-1468
128. Mulligan RC. The basic science of gene therapy. Science, 1993, 260 (5110): 926-932
129. Guilherme N.M, Ferreira Gabriel A, Monterio, et al. Downstream processing of plasmid DNA for gene therapy and DNA vaccine applications. Tibech, 2001,18 (9): 380-388
130. Mor G., Klinman D.M., Shapiro S., et al. Complexity for the cytokine and antibody response elicited by immunizing mice with Plasmodium yoelii circumsporozoite protein plasmid DNA. J. Immunol, 1995, 155 (4): 2039-46
131. Moore AC, Hill. Progress in DNA-based heterologous prime-boost immunization strategies for malaria. Immunological Reviews, 2004, 199: 126-43
132. Sechi L A, Mara L, Cappai P, et al. Immunization with DNA vaccines encoding different mycobacterial antigens elicits a Th1 type immune response in lambs and protects against Mycobacterium avium subspecies paratuberculosis infection. Vaccine, 2006, 24 (3): 229-235
133. Martin T., Parker S.E., Hedstrom R., et al. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Human Gene Therapy, 1999, 10 (5): 759-768
134. Mac Gregor R. R., Boyer J. D., Ciccarelli R.B., et al. Safety and immune responses to a DNA-based human immunodeficiency virus (HIV) type I env/rev vaccine in HIV-infectedrecipients: follow-up data. J. Infect Dis., 2000, 181 (1): 406-412
135. Kanellos T., Sylvester I.D., Ambali A.G. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Human Gene Therapy, 1999, 10 (5): 759-768
136. Martin T., Parker S.E., Hedstrom R., et al. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Human gene therapy, 1999, 10 (5): 759-768
1. Kowalczyk D.W and Ertl H.C.J. Immune responses to DNA vaccines. Cell and Molecular Life Sciences, 1999, 55 (5):751-770
2. Sahota S S, Townsend M and Stevenson F K. Identification and assembly of V genes as idiotype-specific DNA fusion vaccines in multiple myeloma. Methods Mol Med, 2005, 113: 105-19
3. Rosati M, Von Gegerfelt A, Roth P, et al. DNA vaccines expressing different forms of simian immunodeficiency virus antigens decrease viremia upon SIVmac251 challenge. J Virol, 2005, 79 (13): 8480-92
4. Prazeres D.M.F. Large-scale production of pharmaceutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Trends in Biotechnology, 1999, 17 (4): 169-174
5. O’Kennedy R.D, Baldwin C and Keshavarz-Moore E. Effects of growth medium selection on plasmid DNA production and initial processing steps. Journal of Biotechnology, 2000, 76 (8):175-183
6. J.萨姆布鲁克, D.W 拉塞尔. 分子克隆实验指南第三版. 科学出版社
7. Wegrzyn G. Replication of plasmids during bacterial response to amino acid starvation. Plasmid, 1999, 41(1): 1-16
8. Kristala J. P, Sangeetha S, Jason M, et.al. Industrial scale production of plasmid DNA for vaccine and gene therapy: plasmid design, production, and purification. Enzyme and Microbial Technology, 2003, 16 (33): 865–883
1. W Chen. Automated fed-batch fermentation with feed-back controls based on dissolved oxygen (DO) and pH for prouuction of DNA vaccines. Journal of Industrial Microbiology & Biotechnology, 1997, 18 (1): 43-48
2. J.萨姆布鲁克, D.W. 拉塞尔. 分子克隆第三版
3. R.O’Kennedy C.J, Houghton. Effects of growth environment on recombinant plasmid stabilityin Saccharomyces cersvisiae grown in continuous culture. Appl Microbiol Biotechnol, 1995, 44 (1-2): 126-132
4. Corchero J L, Villaverde A. Plasmid maintenance in Escherichia coli recombinant cultures is dra, steadily, and specifically influenced by features of the encoded proteins. Biotechnology and Bioengineering, 1998, 58 (6): 625-632
5. Lahijani R, Hulley G. High-yield production of PBR322-derived plasmids intended for human gene therapy by employing a temperature controllable point mutation. Human Gene Therapy, 1996, 7 (16): 1971-80
6. D.J.Korz. Simple fed-batch technique for high cell density cultivation of Escherichia coli. Journal of Biotechnology, 1995, 39 (1): 59-65
7. Reinikainen P, Korpela K. Escherichia coli production in fermentor. Biotechnology and Bioengineering, 1989, 33: 386–93
8. Kristala Jones Prather. Industrial scale production of plasmid DNA for vaccine and gene therapy: plasmid design, production, and purification. Enzyme and Microbial Technology, 2003, 33 (7): 865-883
1. 荣晓花, 凌沛学. 溶菌酶的研究进展. 中国生化药物杂志, 1999, 20 (6): 916-919
2. 史萸芳. 溶菌酶抑菌作用的实验. 牙体牙髓牙周病学杂志, 1996, 6 (1): 26-28
3. 防治奶牛乳腺炎的基因药物. 专利号:ZL02148545.3
4. Sambrook J, Russell DW. Molecular Cloning A Laboratory Manual. Third Edition, New York: Cold Spring Harbor Laboratory Press, 2002
5. 柳 湘, 田培坤, 顾键人, 等. 非病毒载体系统在基因治疗中的应用. 国外医学肿瘤学分册, 2001, 28 (3): 189-191
6. Kristala Jones Prather, Sangeetha Sagar, Jason Murphy, et al. Industrial scale production of plasmid DNA for vaccine and gene therapy: plasmid design, production, and purification. Enzyme and Microbial Technology, 2003, 33 (7): 865-883
7. Ferber D. Gene therapy: safer and virus free. Science, 2001, 294 (5547): 1638-42
8. Guilherme N.M. Ferreira, Gabriel A. Monteiro, Duarte M.F, et al. Downstream processing of plasmid DNA for gene therapy and DNA vaccine applications. Trends Biotechnol, 2000, 18 (9): 380-388
9. J.Zhang R. Greasham. Chemically defined media for commercial fermentation. Appl Microbiol Biotechnol, 1999, 51: 407-421
10. 陈天寿. 微生物培养基的制造与应用. 中国农业出版社, 1995
1. 薛方明. 防治奶牛乳腺炎基因药物的临床试验与安全平均. 扬州大学学位论文, 2005
2. Birnboim, H.C. and Doly. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res, 1979, 7 (6): 1513-23
1. DuarteM.F.Prazeres, Guilherme N. M. Ferreira, GabrielA. Monteiro, et al. Large-scale production of pharmaeutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Tibtech Appril, 1999, 19 (2): 169-174
2. Duarte Miguel F. Prazeres, Thomas Schluep, Charles Cooney, et al. Preparative purification of supercoiled plasmid DNA using anionexchange chromatography. Journal of Chromatography A, 1998, 806 (1): 31- 45
3. Tseng WC, Ho FL. Enhanced purification of plasmid DNA using Q-Sepharose by modulationof alcohol concentrations. J Chromatogr B Analyt Technol Biomed Life Sci, 2003, 791(1-2): 263-72
4. Eon-Duval A, Burke G. Purification of pharmaceutical-grade plasmid DNA by anion-exchange chromatography in an RNase-free process. J Chromatogr B Analyt Technol Biomed Life Sci, 2004, 804 (2): 327-35
5. Darby RA, Hine AV. LacI-mediated sequence-specific affinity purification of plasmid DNA for therapeutic applications. FASEB J, 2005, 19 (7): 801-3
6. Wahlund PO, Gustavsson PE, Izumrudov VA, et al. Precipitation by polycation as capture step in purification of plasmid DNA from a clarified lysate. Biotechnol Bioeng, 2004, 87 (5): 675-84
7. Shenoy SR, Khalid M, Gupta A, et al. Purification of an Enterobacter aerogenes plasmid DNA using MnCl2 as compaction agent. Anal Biochem, 2003, 321 (2): 256-8
8. Balan S, Murphy J, Galaev I, et al. Metal chelate affinity precipitation of RNA and purification of plasmid DNA. Biotechnol Lett, 2003, 25 (13): 1111-6
9. J.萨姆布鲁克, D.W 拉塞尔, 分子克隆实验指南三版, 科学出版社, 2003
10. M. Susana Levy, Ronan D. O’Kennedy, Parviz Ayazi-Shamlou, et al. Biochemical engineering approaches to the challenges of producing pure plasmid DNA. Trends Biotechnol, 2000, 18 (7): 296-305
11. Alex Eon-Duval, RobertH.MacDuff, Carol A.Fisher, et al. Removal of RNA impurities by tangential flow filtration in an RNase-free plasmid DNA purification process. Analytical Biochemistry, 2003, 316 (2): 66-73
12. Eon-Duval, Gumbs K, Ellett C. Precipitation of RNA impurities with high salt in a plasmid DNA purification process: use of experimental design to determine reaction conditions. Biotechnol Bioeng, 2003, 83 (5): 544-53
13. M.S. Levy, I.J. Collins, J.T. Tsai P., et al. Removal of contaminant nucleic acids by nitrocellulose filtration during pharmaceutical-grade plasmid DNA processing. Journal of Biotechnology, 2000, 76 (2-3): 197-205
14. Lander RJ, Winters MA, Meacle FJ, et al. Fractional precipitation of plasmid DNA from lysateby CTAB. Biotechnol Bioeng, 2002, 79 (7): 776-84
15. Teeters MA, Conrardy SE, Thomas BL. Adsorptive membrane chromatography for purification of plasmid DNA. J Chromatogr A, 2003, 989 (1): 165-73
16. Anspach FB. Endotoxin removal by affinity sorbents. J Biochem Biophys Methods, 2001, 49 (1-3): 665-81
1. D. W. Kowalczyk, H. C. Ertl. Immune response to DNA vaccines. Cellular and Molecular Life Science, 1999, 55: 751-770
2. Robertson J, Griffiths E. WHO Guidelines for Assuring the Quality of DNA Vaccines, Biologicals, 1998, 26 (3): 205-212
3. Guilherme N.M.Ferrira, Gabriel A. Monteiro, Duarte M.F, et al. Downstream processing of plasmid DNA for gene therapy and DNA vaccine applications. Trends Biotechnol, 2000, 18 (9): 380-388
4. May CM, Laurent C, Jean M. A novel potent strategy for gene delivery using a single peptide vector as a carrier. Nucleic Acids Research, 1999, 27 (17): 3510-3517
5. Schuster M, Wu GY, Walton CM. Multicoponent DNA carrier with a vesicular stomatitis virus G-peptide greatly enhances liver targeted gene expression in mice. Bioconjug Chem, 1999, 10 (6): 1075-1083
6. QIAGEN Plasmid Purification Handbook, 2003: 72
7. Martich GD, Boujoukos AJ, Suffredini AF. Response of man to endotoxin. Immunobiology, 1993, 187 (3-5): 403-16
8. Bragonzi A, Boletta A, Motexs A, et al. Comparison between cationic polymers and lipids in mediating systemic gene delivery to the drugs. Gene Ther, 1999, 6 (12): 1995-2004.
9. 王风阳, 孟庆文, 扈容良, 等. 含 LacZ 重组逆转录病毒的制备及其在 NIH3T3 细胞的表达. 中国预防兽医学报, 2001, 23 (5): 329-331
10. 钱锋, 肖成祖. 影响非洲绿猴肾细胞脂质体转染效率的因素. 生物技术通报, 1998, 5: 31-35
11. 廖文俊, 刘彦仿, 刘玉峰. β-半乳糖苷酶基因转染角质形成细胞的瞬时表达检测. 临床皮肤科杂志, 1999, 28 (2): 71-74
12. Kichler A,Leborgne C, Coeytauxe, et al. Polyethylenimine-mediated gene delivery: a mechanistic study. J Gene Med, 2001, 3 (2): 135-144
13. Koping-Hoggard M, Tubuleka I, Guan H, et al. Chitosan as a nonviral gene delivery system structure-property relationships and characteristics compared with polyethylenimine in vitro and after lung administration in vivo. Gene Ther, 2001, 8 (14): 1108-1112
14. Zaumer W, Brunner S, Buschle M, et al. Differential behavior of lipid based and polycation based gene transfer systems in transfecting primary human fibroblasts a potentianl role of polylysine in nuclear transport. Biochim Biophys Acta, 1999, 1428 (1): 57-61
15. Durocher Y, Perret S, Kamen A. High-level and high-throughout recombinant protein production by transient transfection of suspension growing human 293-EBNA1 cells. Nucleic Acids Research, 2002, 30: 1-9
16. 莫国玉, 陈光明, 黄芝瑛, 等. 治疗型 HBV DNA 疫苗在小鼠组织中的分布和长期毒性研究. 解放军医学杂志, 2003, 28 (6): 504-507
17. 李经忠, 王青青. 非病毒载体的研究现状. 国外医学分子生物学分册, 2002, 24 (5): 317-320
18. Ferreira. Plasmid manufacturing-an overview. In Plasmids for Therapy and Vaccination, Schleef M (ed). Whiley-VCH: Weinheim, 2001, 203-204
19. Raetz CR. Biochemistry of endotoxins. Annu Rev Biochem, 1990, 59: 129-170
20. Kastowsky M, Gutberler T, Bradaczek H. Molecular modeling of the three-dimensional structure and conformational flexibility of bacterial lipopolysaccharide. J Bacteriol, 1992,174: 4798-806
1. 莫国玉, 陈光明, 黄芝瑛, 等. 治疗型 HBV DNA 疫苗在小鼠组织中的分布和长期毒性研究. 解放军医学杂志, 2003, 28 (6): 504-507
2. John Donnelly, Karin Berry, Jeffrey B. Ulmer. Technical and regulatory hurdles for DNA vaccines. International Journal for Parasitology, 2003, 33 (5-6): 457-467
3. Kowakczyk DW and Ertl HC. Immune responses to DNA vaccines. Cellular and Molecular Life Sciences, 1999, 55 (5): 751-770
4. Tang D. C., Devit M. and Johnston S. A. Genetic immunization is a simple method for eliciting an immune response. Nature, 1992, 356 (6365): 152-154
5. Prazeres D.M.F. Large-scale production of pharmaceutical-grade plasmid DNA for gene therapy: problems and bottlenecks. Trends in Biotechnology, 1999, 17 (4): 169-174
6. QIAGEN Plasmid Purification Handbook, 2003
7. Robertson J, Griffiths E. WHO Guidelines for Assuring the Quality of DNA Vaccines. Biologicals, 1998, 26 (3): 205-212
8. Dagmar Petsch, Friedrich Birger Anspach. Endotoxin removal from protein solutions. Journal of Biotechnology, 2000, 76 (2-3): 97-119
9. Lydia A. Falk, Leslie K. Ball. Current status and future trends in vaccine regulation-USA. Vaccine, 2001, 19 (13-14): 1567-1572
10. J.萨姆布鲁克, D.W 拉塞尔, 分子克隆实验指南第三版, 科学出版社
1. Verfailliea T, Cox B.M. Goddeeris Immunostimulatory capacity of DNA vaccine vectors in porcine PBMC: a specific role for CpG-motifs. Veterinary Immunology and Immunopathology, 2005, 103 (1-2): 141-151
2. Klinman D M. CpG DNA as a vaccine adjuvant. Expert Rev Vaccines, 2003, 2 (2): 305-15
3. 莫国玉, 陈光明, 黄芝瑛, 等. 治疗型HBV DNA疫苗在小鼠组织中的分布和长期毒性研究. 解放军医学杂志, 2003, 28 (6): 504-507
4. Guidance for Industry Considerations for Plasmid DNA Vaccines for Infectious Disease Indications
5. Wang Z, Troilo P J, Wang X, et al. Detection of integration of plasmid DNA into host genomic DNA following intramuscular injection and electroporation. Gene Therapy, 2004, 11 (8): 711-21
6. Ledwith B J, Manam S, Troilo P J, et al. Plasmid DNA vaccines: assay for integration into host genomic DNA. Dev Biol (Basel), 2000, 104: 33-43
7. Liu Y, Liggitt D, Zhong W, et al. Cationic liposome-mediated intravenous gene delivery. Biol Chem, 1995, 270 (42): 24864-70
8. Mahato K, Kawabata Y, Takakura M, et al. In vivo disposition characteristics of plasmid DNA complexed with cationic liposomes. J Drug Target, 1995, 3 (2): 149-157
9. McLean J.W, Fox E A, Baluk P, et al. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am J Physiol, 1997, 273: H387-404
10. Delphine Lechardeura A.S, Verkmanb and Gergely L. Lukacs. Intracellular routing of plasmid DNA during non-viral gene transfer. Advanced Drug Delivery Reviews, 2005, 57 (5): 755-767
11. Nichols WW, Ledwith BJ, Manam SV, et al. Potential DNA vaccine integration into host cell genome. Ann Nei York Acad Sci, 1995, 772: 30-39