TMEM66基因功能初步研究
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摘要
免疫系统是机体中一个极为重要的防御系统，其功能不正常，机体的生命就会受到威胁。免疫系统中起核心作用的是免疫细胞，均起源于骨髓中的造血干细胞。因此，造血干细胞的正常分化对维持机体免疫系统的功能是至关重要的。TMEM66(transmembrane protein 66)基因可能与免疫细胞的分化密切相关。本研究拟从分子水平、细胞水平及个体水平探讨TMEM66基因的功能。主要研究工作和结果如下：
1 TMEM66基因功能预测
TMEM66基因是一个新的功能基因，目前还没有该基因的专门研究报道，其功能也不清楚。本研究拟通过对TMEM66基本生物学信息进行收集整理，并对TMEM66的结构和功能进行初步预测，为研究该基因的功能奠定一定的基础。结果显示：TMEM66是一个三跨膜蛋白，含有一个DUF1183功能域；人TMEM66基因在分化成熟的免疫细胞如B淋巴细胞、T淋巴细胞、NK细胞、单核细胞、树突细胞中高表达，在脑、脊髓、淋巴结和血液中也高表达，而在白血病细胞(HL60、Molt4、K562)及恶性淋巴瘤细胞系(Raji、Daudi)等未分化成熟的免疫细胞中低表达；在癌变组织中该基因表达下调。以上研究结果表明：TMEM66基因可能与免疫细胞的分化相关，低表达TMEM66基因与癌症发生相关。
2 TMEM66基因分子水平研究
根据生物信息学分析的结果，推测TMEM66基因是一个与细胞分化相关的新基因。本研究拟对猪TMEM66基因进行克隆、定位、多态分析，并对猪和人TMEM66基因的表达规律进行研究，为阐明该基因的功能提供分子生物学依据。结果显示：扩增得到猪TMEM66基因片段长度为1886bp，包含一个完整ORF编码337氨基酸，与人TMEM66蛋白(NP_057211)同源性达84％；猪TMEM66基因定位于猪15号染色体q臂与微卫星SW1673紧密连锁；在猪TMEM66基因CDs区发现了一个A／C多态，基因频率检测发现只在巴马猪群中存在等位基因A且A等位基因频率远远高于C等位基因，其它群体中都只有等位基因C，没有A等位基因；猪TMEM66在肺、脾、淋巴结、脑组织中高表达，而在骨骼肌、心肌和脂肪中低表达；人TMEM66在癌变组织或细胞中比其对应的正常组织表达量下降；TMEM66基因随着胚胎的生长发育呈现上调表达趋势；ATRA诱导U937分化过程中，TMEM66基因呈现上调趋势，48h表达到达高峰，以后呈现下调趋势；以上研究结果表明：TMEM66基因可能与细胞分化相关。
3 TMEM66基因细胞水平研究
TMEM66基因是一个膜蛋白，其亚细胞定位及表达水平与功能密切相关。本研究对人TMEM66基因进行了亚细胞定位；并构建慢病毒过表达载体，研究过表达该基因对细胞周期的影响，进一步从细胞水平探讨TMEM66基因的功能。结果显示：TMEM66分布于细胞核膜上。本研究中发现人TMEM66还存在其它两个转录本。第二转录本比TMEM66缺少了第一跨膜域及部分第一膜内域；第三转录本缺少了第一膜内域及第一膜内域。推测它们可能与TMEM66的功能调节相关。对TMEM66第三个转录本进行亚细胞定位，结果表明第三个转录本分布于细胞质及细胞核膜；对TMEM66互作蛋白IFIT5进行亚细胞定位分析，结果发现IFIT5分布于细胞质中；通过慢病毒介导TMEM66在U937细胞中过表达研究，发现过表达TMEM66导致细胞凋亡增加。经ATRA诱导不同时间点检测细胞周期，结果发现TMEM66过表达的U937细胞G2期比率比对照组高，凋亡比率也比对照组高。以上研究结果表明：TMEM66基因是一个核膜蛋白，过表达TMEM66基因导致细胞凋亡增加，在诱导细胞分化过程中，过表达TMEM66可导致细胞G2期抑制并促使G2期抑制的细胞发生凋亡。
4 TMEM66基因突变体转基因小鼠构建
为了从个体水平探讨TMEM66基因的功能，本研究采用显微注射法制作TMEM66突变体转基因小鼠，为进一步研究该基因的功能奠定基础。结果显示：出生的55只小鼠中，通过PCR法和Southern杂交检测，有6只为阳性鼠，传代研究发现3只阳性鼠可以稳定传代，外源基因片段以串联重复的方式整合到小鼠的基因组中，RT—PCR检测发现转入的突变基因在多个组织中转录。以上研究结果表明：通过显微注射法得到了3只可以传代的转基因阳性鼠，为以后从个体水平研究该基因的功能奠定了基础。
The immune system is the essential part of the body to protect itself to counter detrimental agents from outside. The malfunction of the system sometimes caused lethal problems of normal life. The immune cells play a central role in the system; they derived from hematopoietic progenitor cell in bone marrow. So, the development and differentiation of hematopoietic progenitor cell is vital importance for the normal functioning of immune activities. TMEM66 (transmembrane protein 66) is a protein involved in the differentiation of the immune cells. Present studies focus in the gene function both in vitro and in vivo. The main results are as follows:
1 The predicted function of TMEM66 gene
Until now, there is no report in the scientific literature regarding the function of TEME66 gene. At first, we colleted some related information about this gene employ some bioinformatics methods to deduce the possible biological function. This will be better for future study. The TEME66 protein is a tri-transmembrane protein which contains a DUF1183 domain. In human, the gene expression of TEME66 is high in differentiated immune cells such as B lymphocytes, T lymphocytes, monocytes and dendritic cells. It also showed high expression in brain, spinal cord, lymph note and blood. However, it showed low expression level in leukemic cell (HL60, Molt4 and K562), malignant lymphoma cells (Raji, Daudi) and other undifferentiated immune cells. It is down-regulated in carcinomatous tissues too. These results indicated that TEME66 is involved in the differentiation of immune cells and its aberrant low expression is associated with cancer.
2 Molecular characterization of TEME66 gene
TEME66 is a new gene which in relate to cell differentiation in our bioinformatics predictions. Present study cloned, mapped the porcine TEME66 gene, then identified several polymorphism sites in pig populations and detected its expression pattern in both human and pig, provided basic information for further research of this gene. Our PCR results showed the pig TMEM66 gene contains 1886bp including an integrity ORF and encoding a protein with 337 animo acids which shared 84% homologous with its human counterpart. The pig TMEM66 gene located in the q arm of porcine chromosome 15 and closely linked with marker SW1673. An A/C SNP is identified in the pig TMEM66 locus. The genotyping results showed the A allele is only presented in Bama pigs and it is the dominant allele in this population. However, there is only C allele in other pigs investigated. Pig TEME66 gene expression is abundant in lung, spleen, lymph note and brain, but in low level in skeletal muscle, heart and fat. The expression of human TEME66 gene is down-regulated in carcinomatous tissues or cells compared with normal tissues. The TMEM66 gene is up-regulated through the embryo development. After ATRA induction in U937 cells, the expression of TEME66 gene is increased, reached the plateau around 48 hours and then decreased. These results indicated that TEME66 gene involved in cell differentiation. In this study, there were two other transcriptions of human TMEM66 gene founded. They may participate to the regulation of the function of TMEM66 gene.
3 TEME66 function in cells
TEME66 protein is a membrane protein; the subcellular localization of TEME66 protein could implicate its function. This study detected the subcellular localization of human TEME66 protein. We also studyed the overexpression of TMEM66 gene using lentivirus vector. The results indicated that TEME66 protein located in the nuclear membrane. The third transcript distributed across the cytoplasm and also nuclear membrane. The IFIT5 which was interacted with TMEM66 localized to cytoplasm. After overexpression of TEME66 by lentivirus infection in U937 cells, the apoptosis intensified. Then, ATRA induction was introduced in and the G2 period of U937 cells with overexpression of TEME66 is higher than that of control cells without TEME66 overexpression. The apoptosis percent is also higher in the overexression cell lines. These results indicated that TEME66 protein mainly localized to the nuclear membrane. During differentiation, TEME66 could cause the G2 period prolongation and induce the apoptosis of targeted G2 period cells.
4 The TEME66 mutation transgenic mouse
To investigate the TEME66 function in vivo, we produced the TEME66 mutation transgenic mice by conventional micro injection method. After PCR and southern blot verification, we found 6 positive individuals among the 55 neonatal pups. Three positive mice could stably transfer the transgene to next generation through passage. The transgene is incorporated into the host genome in a serial multi-copy manner. RT-PCR analysis indicated that the transgene was expressed in many tissues. The results showed at least 3 mice embraced the TEME66 mutation transgene which could be stably passaged; provide a valuable model for the further study of TEME66 function.
1 TMEM66基因功能预测
TMEM66基因是一个新的功能基因，目前还没有该基因的专门研究报道，其功能也不清楚。本研究拟通过对TMEM66基本生物学信息进行收集整理，并对TMEM66的结构和功能进行初步预测，为研究该基因的功能奠定一定的基础。结果显示：TMEM66是一个三跨膜蛋白，含有一个DUF1183功能域；人TMEM66基因在分化成熟的免疫细胞如B淋巴细胞、T淋巴细胞、NK细胞、单核细胞、树突细胞中高表达，在脑、脊髓、淋巴结和血液中也高表达，而在白血病细胞(HL60、Molt4、K562)及恶性淋巴瘤细胞系(Raji、Daudi)等未分化成熟的免疫细胞中低表达；在癌变组织中该基因表达下调。以上研究结果表明：TMEM66基因可能与免疫细胞的分化相关，低表达TMEM66基因与癌症发生相关。
2 TMEM66基因分子水平研究
根据生物信息学分析的结果，推测TMEM66基因是一个与细胞分化相关的新基因。本研究拟对猪TMEM66基因进行克隆、定位、多态分析，并对猪和人TMEM66基因的表达规律进行研究，为阐明该基因的功能提供分子生物学依据。结果显示：扩增得到猪TMEM66基因片段长度为1886bp，包含一个完整ORF编码337氨基酸，与人TMEM66蛋白(NP_057211)同源性达84％；猪TMEM66基因定位于猪15号染色体q臂与微卫星SW1673紧密连锁；在猪TMEM66基因CDs区发现了一个A／C多态，基因频率检测发现只在巴马猪群中存在等位基因A且A等位基因频率远远高于C等位基因，其它群体中都只有等位基因C，没有A等位基因；猪TMEM66在肺、脾、淋巴结、脑组织中高表达，而在骨骼肌、心肌和脂肪中低表达；人TMEM66在癌变组织或细胞中比其对应的正常组织表达量下降；TMEM66基因随着胚胎的生长发育呈现上调表达趋势；ATRA诱导U937分化过程中，TMEM66基因呈现上调趋势，48h表达到达高峰，以后呈现下调趋势；以上研究结果表明：TMEM66基因可能与细胞分化相关。
3 TMEM66基因细胞水平研究
TMEM66基因是一个膜蛋白，其亚细胞定位及表达水平与功能密切相关。本研究对人TMEM66基因进行了亚细胞定位；并构建慢病毒过表达载体，研究过表达该基因对细胞周期的影响，进一步从细胞水平探讨TMEM66基因的功能。结果显示：TMEM66分布于细胞核膜上。本研究中发现人TMEM66还存在其它两个转录本。第二转录本比TMEM66缺少了第一跨膜域及部分第一膜内域；第三转录本缺少了第一膜内域及第一膜内域。推测它们可能与TMEM66的功能调节相关。对TMEM66第三个转录本进行亚细胞定位，结果表明第三个转录本分布于细胞质及细胞核膜；对TMEM66互作蛋白IFIT5进行亚细胞定位分析，结果发现IFIT5分布于细胞质中；通过慢病毒介导TMEM66在U937细胞中过表达研究，发现过表达TMEM66导致细胞凋亡增加。经ATRA诱导不同时间点检测细胞周期，结果发现TMEM66过表达的U937细胞G2期比率比对照组高，凋亡比率也比对照组高。以上研究结果表明：TMEM66基因是一个核膜蛋白，过表达TMEM66基因导致细胞凋亡增加，在诱导细胞分化过程中，过表达TMEM66可导致细胞G2期抑制并促使G2期抑制的细胞发生凋亡。
4 TMEM66基因突变体转基因小鼠构建
为了从个体水平探讨TMEM66基因的功能，本研究采用显微注射法制作TMEM66突变体转基因小鼠，为进一步研究该基因的功能奠定基础。结果显示：出生的55只小鼠中，通过PCR法和Southern杂交检测，有6只为阳性鼠，传代研究发现3只阳性鼠可以稳定传代，外源基因片段以串联重复的方式整合到小鼠的基因组中，RT—PCR检测发现转入的突变基因在多个组织中转录。以上研究结果表明：通过显微注射法得到了3只可以传代的转基因阳性鼠，为以后从个体水平研究该基因的功能奠定了基础。
The immune system is the essential part of the body to protect itself to counter detrimental agents from outside. The malfunction of the system sometimes caused lethal problems of normal life. The immune cells play a central role in the system; they derived from hematopoietic progenitor cell in bone marrow. So, the development and differentiation of hematopoietic progenitor cell is vital importance for the normal functioning of immune activities. TMEM66 (transmembrane protein 66) is a protein involved in the differentiation of the immune cells. Present studies focus in the gene function both in vitro and in vivo. The main results are as follows:
1 The predicted function of TMEM66 gene
Until now, there is no report in the scientific literature regarding the function of TEME66 gene. At first, we colleted some related information about this gene employ some bioinformatics methods to deduce the possible biological function. This will be better for future study. The TEME66 protein is a tri-transmembrane protein which contains a DUF1183 domain. In human, the gene expression of TEME66 is high in differentiated immune cells such as B lymphocytes, T lymphocytes, monocytes and dendritic cells. It also showed high expression in brain, spinal cord, lymph note and blood. However, it showed low expression level in leukemic cell (HL60, Molt4 and K562), malignant lymphoma cells (Raji, Daudi) and other undifferentiated immune cells. It is down-regulated in carcinomatous tissues too. These results indicated that TEME66 is involved in the differentiation of immune cells and its aberrant low expression is associated with cancer.
2 Molecular characterization of TEME66 gene
TEME66 is a new gene which in relate to cell differentiation in our bioinformatics predictions. Present study cloned, mapped the porcine TEME66 gene, then identified several polymorphism sites in pig populations and detected its expression pattern in both human and pig, provided basic information for further research of this gene. Our PCR results showed the pig TMEM66 gene contains 1886bp including an integrity ORF and encoding a protein with 337 animo acids which shared 84% homologous with its human counterpart. The pig TMEM66 gene located in the q arm of porcine chromosome 15 and closely linked with marker SW1673. An A/C SNP is identified in the pig TMEM66 locus. The genotyping results showed the A allele is only presented in Bama pigs and it is the dominant allele in this population. However, there is only C allele in other pigs investigated. Pig TEME66 gene expression is abundant in lung, spleen, lymph note and brain, but in low level in skeletal muscle, heart and fat. The expression of human TEME66 gene is down-regulated in carcinomatous tissues or cells compared with normal tissues. The TMEM66 gene is up-regulated through the embryo development. After ATRA induction in U937 cells, the expression of TEME66 gene is increased, reached the plateau around 48 hours and then decreased. These results indicated that TEME66 gene involved in cell differentiation. In this study, there were two other transcriptions of human TMEM66 gene founded. They may participate to the regulation of the function of TMEM66 gene.
3 TEME66 function in cells
TEME66 protein is a membrane protein; the subcellular localization of TEME66 protein could implicate its function. This study detected the subcellular localization of human TEME66 protein. We also studyed the overexpression of TMEM66 gene using lentivirus vector. The results indicated that TEME66 protein located in the nuclear membrane. The third transcript distributed across the cytoplasm and also nuclear membrane. The IFIT5 which was interacted with TMEM66 localized to cytoplasm. After overexpression of TEME66 by lentivirus infection in U937 cells, the apoptosis intensified. Then, ATRA induction was introduced in and the G2 period of U937 cells with overexpression of TEME66 is higher than that of control cells without TEME66 overexpression. The apoptosis percent is also higher in the overexression cell lines. These results indicated that TEME66 protein mainly localized to the nuclear membrane. During differentiation, TEME66 could cause the G2 period prolongation and induce the apoptosis of targeted G2 period cells.
4 The TEME66 mutation transgenic mouse
To investigate the TEME66 function in vivo, we produced the TEME66 mutation transgenic mice by conventional micro injection method. After PCR and southern blot verification, we found 6 positive individuals among the 55 neonatal pups. Three positive mice could stably transfer the transgene to next generation through passage. The transgene is incorporated into the host genome in a serial multi-copy manner. RT-PCR analysis indicated that the transgene was expressed in many tissues. The results showed at least 3 mice embraced the TEME66 mutation transgene which could be stably passaged; provide a valuable model for the further study of TEME66 function.
引文
1．陈海金，黄宗海，苏国强，吴爱国，俞金龙．慢病毒介导的双自杀基因对大肠癌细胞的靶向杀伤作用．世界华人消化杂志，2006 14(17)：1681．1687
2．陈晓萍，徐作武，姜薇．造血细胞发育调控机制的研究进展．浙江工业大学学报，2007，35(1)：6-9
3．陈竺，强伯勤，方福德．基因组科学与人类疾病．北京：科学出版社，2001
4．高燕，林莉萍，丁健．细胞周期调控的研究进展．生命科学，2005，17(4)：318-322
5．李海燕，尤艳，尤德渊．干扰素治疗肿瘤机制研究的进展．实用肿瘤学杂志，2006， 20(3)：245-246
6．李振宇，徐开林，潘秀英．慢病毒载体构建及结构优化．国外医学分子生物学分册，2002，24(5)： 310-313
7．刘春宇，张春玲，夏家辉．数据综合分析鉴定人类Auxilin基因．生物化学与生物物理研究进展，1998，25(5)：434-438
8．欧阳五庆，李谱华，林成招．细胞周期及其调控研究进展．中国兽医科技，2003，33(2)：35-40
9．庞月华，张新平，冯义伯，何善述．细胞周期及其调控研究进展．新疆医科大学学报，2005，28(6)：596-598
10．孙亚萍，王英明，乔守怡．干扰素及其最新研究进展．中国免疫学杂志，2006，22： 676-679
11．王霞，李萍．细胞周期调控与肿瘤研究进展．中国医学检验杂志，2006，7(1)：59-61
12．吴波，朱作言．转基因动物整合位点的研究进展．遗传HEREDITAS，2003，25(1)：77-80
13．吴楠，查锡良．肿瘤细胞的诱导分化．生命的化学，2004，24(6)：496-498
14．吴艳萍，李立平，张颖，刘嘉琳，高伟敏．细胞周期调控分子与肿瘤关系的研究进展．军医进修学院学报，2006，27(4)：306-308
15．许培权，龚建平．全反式维甲酸诱导HL60细胞分化的细胞周期时相性研究．华中科技大学学报(医学板)，2004，33(4)：445-448
16 赵卫红, 寿好长, 闰福岭. 细胞凋亡. 郑州: 河南医科大学出版社, 1997, 7-11.
17. Aboul-ela F, Karn J, Varani, G. The structure of the human immunodeficiency virus type-1 TAR RNA reveals principles of RNA recognition by Tat protein. J Mol Biol, 1995, 253:313-32
18. Azzouz M, Martin-Rendon E, Barber R D, Mitrophanous K A, Carter E E, Rohll J B, Kingsman S M, Kingsman A J, Mazarakis N D. Multicistronic lentiviral vector-mediated striatal gene transfer of aromatic L-amino acid decarboxylase, tyrosine hydroxylase, and GTP cyclohydrolase I induces sustained transgene expression, dopamine production, and functional improvement in a rat model of Parkinson's disease. J Neurosci, 2002, 22:10302-10312
19. Barinaga M. Is apoptosis key in Alzheimer's disease? Science, 1998,281:1303-1304
20. Bensadoun J C, Deglon N, Tseng J L, Ridet J L, Zurn A D, Aebischer P. Lentiviral vectors as a gene delivery system in the mouse midbrain: cellular and behavioral improvements in a 6-OHDA model of Parkinson's disease using GDNF. Exp Neurol, 2000,164: 15-24
21. Bishop J O, Smith P. Mechanism of chromosomal integration of microinjected DNA. Mol Biol Med, 1989, 6: 283-298
22. Brinster R L, Avarbock M R. Germline transmission of donor haplotype following spermatogonial transplantation. Proc Natl Acad Sci U S A, 1994,91:11303-11307
23. Brinster R L, Zimmermann J W. Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci U S A, 1994, 91:11298-11302
24. Buske C, Feuring-Buske M, Abramovich C, Spiekermann K, Eaves C J, Coulombel L, Sauvageau G, Hogge D E, Humphries R K. Deregulated expression of HOXB4 enhances the primitive growth activity of human hematopoietic cells. Blood,2002, 100: 862-868
25. Chang L J, Zaiss A K.Self-inactivating lentiviral vectors and a sensitive Cre-loxP reporter system. Methods Mol Med,2003, 76: 367-382
26. Chen S, Le W. Neuroprotective therapy in Parkinson disease. Am J Ther, 2006, 13: 445-457
27. Clapham P R, Weiss R A. Immunodeficiency viruses. Spoilt for choice of co-receptors. Nature,1997, 388: 230-231
28. Cohen E A, Subbramanian R A, Gottlinger H G. Role of auxiliary proteins in retroviral morphogenesis. Curr Top Microbiol Immunol,1996, 214: 219-235
29. Colombo S, Longhi R, Alcaro S, Ortuso F, Sprocati T, Flora A, Borgese N. N-myristoylation determines dual targeting of mammalian NADH-cytochrome b5 reductase to ER and mitochondrial outer membranes by a mechanism of kinetic partitioning. J Cell Biol, 2005,168: 735-745
30. Daoud R, Mies G, Smialowska A, Olah L, Hossmann KA, Stamm S. Ischemia induces a translocation of the splicing factor tra2-beta 1 and changes alternative splicing patterns in the brain. J Neurosci, 2002, 22: 5889-5899
31. Fire A, Xu S, Montgomery M K, Kostas S A, Driver S E, Mello C C. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 1998, 391: 806-811
32. Fleury C, Mignotte B. Vayssiere J L. Mitochondrial reactive oxygen species in cell death signaling. Biochimie, 2002, 84:131-141
33. Forrest M S, Lan Q, Hubbard A E, Zhang L, Vermeulen R, Zhao X, Li G, Wu Y Y, Shen M, Yin S, Chanock S J, Rothman N, Smith M T. Discovery of novel biomarkers by microarray analysis of peripheral blood mononuclear cell gene expression in benzene-exposed workers. Environ Health Perspect,2005,113: 801-807
34. Frankel A D, Young J A. HIV-1: fifteen proteins and an RNA. Annu Rev Biochem, 1998, 67: 1-25
35. Gerolami R, Uch R, Faivre J, Garcia S, Hardwigsen J, Cardoso J, Mathieu S, Bagnis C, Brechot C, Mannoni P. Herpes simplex virus thymidine kinase-mediated suicide gene therapy for hepatocellular carcinoma using HIV-1-derived lentiviral vectors. J Hepatol, 2004, 40: 291-297
36. Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H. Transcriptional activation by tetracyclines in mammalian cells. Science,1995, 268: 1766-1769
37. Griffith T S, Rauch C T, Smolak P J, Waugh J Y, Boiani N, Lynch D H, Smith C A, Goodwin R G, Kubin M Z. Functional analysis of TRAIL receptors using monoclonal antibodies. J Immunol, 1999,162: 2597-2605
38. Gutierrez-Adan A, Pintado B. Effect of flanking matrix attachment regions on the expression of microinjected transgenes during preimplantation development of mouse embryos. Transgenic Res, 2000, 9: 81-89
39. Hamada T, Sasaki H, Seki R, Sakaki Y. Mechanism of chromosomal integration of transgenes in microinjected mouse eggs: sequence analysis of genome-transgene and transgene-transgene junctions at two loci. Gene, 1993,128: 197-202
40. Hamra F K, Gatlin J, Chapman K M, Grellhesl D M, Garcia J V, Hammer R E, Garbers D L. Production of transgenic rats by lentiviral transduction of male germ-line stem cells. Proc Natl Acad Sci U S A,2002, 99:14931-14936
41. Hanamura A, Caceres J F, Mayeda A, Franza B R, Jr, Krainer A R. Regulated tissue-specific expression of antagonistic pre-mRNA splicing factors. Rna, 1998, 4: 430-444
42. Harrison P M, Kumar A, Lang N, Snyder M, Gerstein M. A question of size: the eukaryotic proteome and the problems in defining it. Nucleic Acids Res, 2002, 30: 1083-1090
43. Hawken R J, Murtaugh J, Flickinger G H, Yerle M, Robic A, Milan D, Gellin J, Beattie C W, Schook L B, Alexander LJ. A first-generation porcine whole-genome radiation hybrid map. Mamm Genome, 1999,10: 824-830
44. Hernandez L D, Hoffman L R, Wolfsberg T G, White J M. Virus-cell and cell-cell fusion. Annu Rev Cell Dev Biol, 1996,12: 627-661
45. Hofmann A, Kessler B, Ewerling S, Weppert M, Vogg B, Ludwig H, Stojkovic M, Boelhauve M, Brem G, Wolf E, Pfeifer A. Efficient transgenesis in farm animals by lentiviral vectors. EMBO Rep,2003, 4: 1054-1060
46. Honaramooz, A, Behboodi E, Blash S, Megee S O, Dobrinski I. Germ cell transplantation in goats. Mol Reprod Dev,2003, 64: 422-428
47. Honaramooz A, Megee S O, Dobrinski I. Germ cell transplantation in pigs. Biol Reprod, 2002, 66: 21-28
48. Indraccolo S, Habeler W, Tisato V, Stievano L, Piovan E, Tosello V, Esposito G, Wagner R, Uberla K, Chieco-Bianchi L, Amadori A. Gene transfer in ovarian cancer cells: a comparison between retroviral and lentiviral vectors. Cancer Res,2002, 62: 6099-6107
49. Indraccolo S, Tisato V, Tosello V, Habeler W, Esposito G, Moserle L, Stievano L, Persano L, Chieco-Bianchi L, Amadori A. Interferon-alpha gene therapy by lentiviral vectors contrasts ovarian cancer growth through angiogenesis inhibition. Hum Gene Ther, 2005,16: 957-970
50. Izadyar F, Den Ouden K, Stout T A, Stout J, Coret J, Lankveld D P, Spoormakers T J, Colenbrander B, Oldenbroek JK, Van der Ploeg K D, Woelders H, Kal H B, De Rooij D G. Autologous and homologous transplantation of bovine spermatogonial stem cells. Reproduction, 2003,126: 765-774
51. Jaenisch R. Germ line integration and Mendelian transmission of the exogenous Moloney leukemia virus. Proc Natl Acad Sci U S A,1976, 73:1260-1264
52. Kafri T, van Praag H, Gage F H, Verma I M. Lentiviral vectors: regulated gene expression. Mol Ther, 2000,1: 516-521
53. Kampa D, Cheng J, Kapranov P, Yamanaka M, Brubaker S, Cawley S, Drenkow J, Piccolboni A, Bekiranov S, Helt G, Tammana H, Gingeras T R. Novel RNAs identified from an in-depth analysis of the transcriptome of human chromosomes 21 and 22. Genome Res, 2004,14: 331-342
54. Kordower J H. In vivo gene delivery of glial cell line-derived neurotrophic factor for Parkinson's disease. Ann Neurol 53 Suppl 3, 2003, S120-132; discussion S132-134
55. Krawczak M, Reiss J, Cooper D N. The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: causes and consequences. Hum Genet, 1992, 90: 41-54
56. Lai Z, Brady R O. Gene transfer into the central nervous system in vivo using a recombinanat lentivirus vector. J Neurosci Res, 2002, 67: 363-371
57. Lamb R A, Pinto L H. Do Vpu and Vpr of human immunodeficiency virus type 1 and NB of influenza B virus have ion channel activities in the viral life cycles? Virology, 1997,229: 1-11
58. Lander E S, Linton L M, Birren B, Nusbaum C, Zody M C, Baldwin J, Devon K, Dewar K, Doyle M, FitzHugh W, Funke R, Gage D, Harris K, Heaford A, Howland J, Kann L, Lehoczky J, LeVine R, McEwan P, McKernan K, Meldrim J, Mesirov J P, Miranda C, Morris W, Naylor J, Raymond C, Rosetti M, Santos R, Sheridan A, Sougnez C, Stange-Thomann N, Stojanovic N, Subramanian A, Wyman D, Rogers J, Sulston J, Ainscough R, Beck S, Bentley D, Burton J, Clee C, Carter N, Coulson A, Deadman R, Deloukas P, Dunham A, Dunham I, Durbin R, French L, Grafham D, Gregory S, Hubbard T, Humphray S, Hunt A, Jones M, Lloyd C, McMurray A, Matthews L, Mercer S, Milne S, Mullikin J C, Mungall A, Plumb R, Ross M, Shownkeen R, Sims S, Waterston R H, Wilson R K, Hillier L W, McPherson J D, Marra M A, Mardis E R, Fulton L A, Chinwalla AT, Pepin K H, Gish W R, Chissoe S L, Wendl M C, Delehaunty K D, Miner T L, Delehaunty A, Kramer J B, Cook L L, Fulton RS, Johnson D L, Minx P J, Clifton S W, Hawkins T, Branscomb E, Predki P, Richardson P, Wenning S, Slezak T, Doggett N, Cheng J F, Olsen A, Lucas S, Elkin C, Uberbacher E, Frazier M, et al. Initial sequencing and analysis of the human genome. Nature, 2001, 409: 860-921
59. Le Gall S, Heard J M, Schwartz O. Analysis of Nef-induced MHC-I endocytosis. Res Virol, 1997,148: 43-47
60. Lois C, Hong E J, Pease S, Brown E J, Baltimore D. Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science, 2002, 295: 868-872
61. Mahe D, Fischer N, Decimo D, Fuchs J P. Spatiotemporal regulation of hnRNP M and 2H9 gene expression during mouse embryonic development. Biochim Biophys Acta, 2000,1492: 414-424
62. Makarova I V, Tarantul V Z, Gazarian KG. [Structural features of the integration site of foreign DNA in the transgenic mouse genome]. Mol Biol (Mosk) , 1998, 22: 1553-1561
63. Mannherz O, Mertens D, Hahn M, Lichter P. Functional screening for proapoptotic genes by reverse transfection cell array technology. Genomics, 2006, 87: 665-672
64. Merilainen O, Hakkarainen T, Wahlfors T, Pellinen R, Wahlfors J. HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct. Int J Oncol, 2005, 27: 203-208
65. Metharom P, Ellem K A, Schmidt C, Wei M Q. Lentiviral vector-mediated tyrosinase-related protein 2 gene transfer to dendritic cells for the therapy of melanoma. Hum Gene Ther, 2001,12: 2203-2213
66. Milan D, Hawken R, Cabau C, Leroux S, Genet C, Lahbib Y, Tosser G, Robic A, Hatey F, Alexander L, Beattie C, Schook L, Yerle M, Gellin J. IMpRH server: an RH mapping server available on the Web. Bioinformatics, 2000,16: 558-559
67. Miyoshi H, Blomer U, Takahashi M, Gage F H, Verma I M. Development of a self-inactivating lentivirus vector. J Virol, 1998, 72: 8150-8157
68. Modrek B, Lee C. A genomic view of alternative splicing. Nat Genet, 2002, 30: 13-19
69. Morgan A, Burgoyne R D, Barclay J W, Craig T J, Prescott G R, Ciufo L F, Evans G J. Graham M E. Regulation of exocytosis by protein kinase C. Biochem Soc Trans,2005, 33:1341-1344
70. Nagano M, Brinster C J, Orwig K E, Ryu B Y, Avarbock M R, Brinster R L. Transgenic mice produced by retroviral transduction of male germ-line stem cells. Proc Natl Acad Sci U S A, 2001, 98:13090-13095
71. Nagano M, Shinohara T, Avarbock M R, Brinster R L. Retrovirus-mediated gene delivery into male germ line stem cells. FEBS Lett, 2000, 475: 7-10
72. Nakai K, Sakamoto H. Construction of a novel database containing aberrant splicing mutations of mammalian genes. Gene, 1994,141: 171-177
73. Naldini L, Blomer U, Gage F H, Trono D, Verma I M. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci U S A, 1996, 93: 11382-11388
74. Palmiter R D, Brinster R L. Transgenic mice. Cell, 1985, 41: 343-345
75. Pan G, O'Rourke K, Chinnaiyan A M, Gentz R, Ebner R, Ni J, Dixit V M. The receptor for the cytotoxic ligand TRAIL. Science, 1997, 276: 111-113
76. Park F, Ohashi K, Kay M A. Therapeutic levels of human factor VIII and IX using HIV-1-based lentiviral vectors in mouse liver. Blood,2000, 96:1173-1176
77. Pawliuk R, Westerman K A, Fabry M E, Payen E, Tighe R, Bouhassira E E, Acharya S A, Ellis J, London I M, Eaves C J, Humphries R K, Beuzard Y, Nagel R L, Leboulch P. Correction of sickle cell disease in transgenic mouse models by gene therapy. Science, 2001, 294: 2368-2371
78. Pfeifer A, Ikawa M, Dayn Y, Verma I M. Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proc Natl Acad Sci U S A, 2002, 99: 2140-2145
79. Ponder K P. Gene therapy for hemophilia. Curr Opin Hematol, 2006,13: 301-307
80. Puglisi J D, Tan R, Calnan B J, Frankel A D, Williamson JR. Conformation of the TAR RNA-arginine complex by NMR spectroscopy. Science, 1992, 257: 76-80
81. Reiser J, Lai Z, Zhang X Y, Brady R O. Development of multigene and regulated lentivirus vectors. J Virol, 2000, 74:10589-10599
82. Ribas A. Genetically modified dendritic cells for cancer immunotherapy. Curr Gene Ther, 2005, 5: 619-628
83. Rinn J L, Euskirchen G, Bertone P, Martone R, Luscombe N M, Hartman S, Harrison P M, Nelson F K, Miller P, Gerstein M, Weissman S, Snyder M. The transcriptional activity of human Chromosome 22. Genes Dev, 2003,17: 529-540
84. Rozovskaia T, Feinstein E, Mor O, Foa R, Blechman J, Nakamura T, Croce C M, Cimino G. Canaani E. Upregulation of Meisl and HoxA9 in acute lymphocytic leukemias with the t (4 :11) abnormality. Oncogene, 2001, 20: 874-878
85. Russo I H, Russo J. Hormonal approach to breast cancer prevention. J Cell Biochem Suppl, 2000, 34:1-6
86. Scorilas A, Kyriakopoulou L, Katsaros D, Diamandis E P. Cloning of a gene (SR-A1) , encoding for a new member of the human Ser/Arg-rich family of pre-mRNA splicing factors: overexpression in aggressive ovarian cancer. Br J Cancer, 2001, 85: 190-198
87. Song Y, Zhang Z, Yu X, Yan M, Zhang X, Gu S, Stuart T, Liu C, Reiser J, Zhang Y, Chen Y. Application of lentivirus-mediated RNAi in studying gene function in mammalian tooth development. Dev Dyn, 2006, 235:1334-1344
88. Stahlberg H, Fotiadis D, Scheuring S, Remigy H, Braun T, Mitsuoka K, Fujiyoshi Y, Engel A. Two-dimensional crystals: a powerful approach to assess structure, function and dynamics of membrane proteins. FEBS Lett, 2001, 504:166-172
89. Stamm S. Signals and their transduction pathways regulating alternative splicing: a new dimension of the human genome. Hum Mol Genet, 2002,11: 2409-2416
90. Stelzl U, Worm U, Lalowski M, Haenig C, Brembeck F H, Goehler H, Stroedicke M, Zenkner M, Schoenherr A, Koeppen S, Timm J, Mintzlaff S, Abraham C, Bock N, Kietzmann S, Goedde A, Toksoz E, Droege A, Krobitsch S, Korn B, Birchmeier W, Lehrach H, Wanker E E. A human protein-protein interaction network: a resource for annotating the proteome. Cell, 2005,122: 957-968
91. Stickeler E, Kittrell F, Medina D, Berget S M. Stage-specific changes in SR splicing factors and alternative splicing in mammary tumorigenesis. Oncogene, 1999, 18: 3574-3582
92. Stutz F, Neville M, Rosbash M. Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast. Cell, 1995, 82: 495-506
93. Sun M, Zhang G R, Kong L, Holmes C, Wang X, Zhang W, Goldstein D S, Geller AI. Correction of a rat model of Parkinson's disease by coexpression of tyrosine hydroxylase and aromatic amino acid decarboxylase from a helper virus-free herpes simplex virus type 1 vector. Hum Gene Ther, 2003,14: 415-424
94. Ullman K S, Powers M A, Forbes D J. Nuclear export receptors: from importin to exportin. Cell, 1997, 90: 967-970
95. Vaux D L. Caspases and apoptosis - biology and terminology. Cell Death Differ, 1999, 6: 493-494
96. Venter J C, Adams M D, Myers E W, Li P W, Mural R J, Sutton G G, Smith H O, Yandell M, Evans C A, Holt R A, Gocayne J D, Amanatides P, Ballew R M, Huson D H, Wortman J R, Zhang Q, Kodira C D, Zheng X H, Chen L, Skupski M, Subramanian G, et al. The sequence of the human genome. Science, 2001, 291: 1304-1351
97. Walczak H, Degli-Esposti M A, Johnson R S, Smolak P J, Waugh J Y, Boiani N, Timour M S, Gerhart M J, Schooley K A, Smith C A, Goodwin R G, Rauch C T. TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL. Embo J, 1997, 16: 5386-5397
98. Yamane K, Kinsella T J. Casein kinase 2 regulates both apoptosis and the cell cycle following DNA damage induced by 6-thioguanine. Clin Cancer Res, 2005, 11: 2355-2363
99. Yerle M, Pinton P, Robic A, Alfonso A, Palvadeau Y, Delcros C, Hawken R, Alexander L, Beattie C, Schook L, Milan D, Gellin J. Construction of a whole-genome radiation hybrid panel for high-resolution gene mapping in pigs. Cytogenet Cell Genet, 1998, 82:182-188
100.Yu X, Zhan X, D'Costa J, Tanavde V M, Ye Z, Peng T, Malehorn M T, Yang X, Civin C I Cheng L. Lentiviral vectors with two independent internal promoters transfer high-level expression of multiple transgenes to human hematopoietic stem-progenitor cells. Mol Ther, 2003, 7: 827-838
101.Zheng J, Kitajima K, Sakai E, Kimura T, Minegishi N, Yamamoto M, Nakano T. Differential effects of GATA-1 on proliferation and differentiation of erythroid lineage cells. Blood, 2006,107: 520-527
102.Zoraqi G, Spadafora C. Integration of foreign DNA sequences into mouse sperm genome. DNA Cell Biol, 1997,16: 291-300
2．陈晓萍，徐作武，姜薇．造血细胞发育调控机制的研究进展．浙江工业大学学报，2007，35(1)：6-9
3．陈竺，强伯勤，方福德．基因组科学与人类疾病．北京：科学出版社，2001
4．高燕，林莉萍，丁健．细胞周期调控的研究进展．生命科学，2005，17(4)：318-322
5．李海燕，尤艳，尤德渊．干扰素治疗肿瘤机制研究的进展．实用肿瘤学杂志，2006， 20(3)：245-246
6．李振宇，徐开林，潘秀英．慢病毒载体构建及结构优化．国外医学分子生物学分册，2002，24(5)： 310-313
7．刘春宇，张春玲，夏家辉．数据综合分析鉴定人类Auxilin基因．生物化学与生物物理研究进展，1998，25(5)：434-438
8．欧阳五庆，李谱华，林成招．细胞周期及其调控研究进展．中国兽医科技，2003，33(2)：35-40
9．庞月华，张新平，冯义伯，何善述．细胞周期及其调控研究进展．新疆医科大学学报，2005，28(6)：596-598
10．孙亚萍，王英明，乔守怡．干扰素及其最新研究进展．中国免疫学杂志，2006，22： 676-679
11．王霞，李萍．细胞周期调控与肿瘤研究进展．中国医学检验杂志，2006，7(1)：59-61
12．吴波，朱作言．转基因动物整合位点的研究进展．遗传HEREDITAS，2003，25(1)：77-80
13．吴楠，查锡良．肿瘤细胞的诱导分化．生命的化学，2004，24(6)：496-498
14．吴艳萍，李立平，张颖，刘嘉琳，高伟敏．细胞周期调控分子与肿瘤关系的研究进展．军医进修学院学报，2006，27(4)：306-308
15．许培权，龚建平．全反式维甲酸诱导HL60细胞分化的细胞周期时相性研究．华中科技大学学报(医学板)，2004，33(4)：445-448
16 赵卫红, 寿好长, 闰福岭. 细胞凋亡. 郑州: 河南医科大学出版社, 1997, 7-11.
17. Aboul-ela F, Karn J, Varani, G. The structure of the human immunodeficiency virus type-1 TAR RNA reveals principles of RNA recognition by Tat protein. J Mol Biol, 1995, 253:313-32
18. Azzouz M, Martin-Rendon E, Barber R D, Mitrophanous K A, Carter E E, Rohll J B, Kingsman S M, Kingsman A J, Mazarakis N D. Multicistronic lentiviral vector-mediated striatal gene transfer of aromatic L-amino acid decarboxylase, tyrosine hydroxylase, and GTP cyclohydrolase I induces sustained transgene expression, dopamine production, and functional improvement in a rat model of Parkinson's disease. J Neurosci, 2002, 22:10302-10312
19. Barinaga M. Is apoptosis key in Alzheimer's disease? Science, 1998,281:1303-1304
20. Bensadoun J C, Deglon N, Tseng J L, Ridet J L, Zurn A D, Aebischer P. Lentiviral vectors as a gene delivery system in the mouse midbrain: cellular and behavioral improvements in a 6-OHDA model of Parkinson's disease using GDNF. Exp Neurol, 2000,164: 15-24
21. Bishop J O, Smith P. Mechanism of chromosomal integration of microinjected DNA. Mol Biol Med, 1989, 6: 283-298
22. Brinster R L, Avarbock M R. Germline transmission of donor haplotype following spermatogonial transplantation. Proc Natl Acad Sci U S A, 1994,91:11303-11307
23. Brinster R L, Zimmermann J W. Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci U S A, 1994, 91:11298-11302
24. Buske C, Feuring-Buske M, Abramovich C, Spiekermann K, Eaves C J, Coulombel L, Sauvageau G, Hogge D E, Humphries R K. Deregulated expression of HOXB4 enhances the primitive growth activity of human hematopoietic cells. Blood,2002, 100: 862-868
25. Chang L J, Zaiss A K.Self-inactivating lentiviral vectors and a sensitive Cre-loxP reporter system. Methods Mol Med,2003, 76: 367-382
26. Chen S, Le W. Neuroprotective therapy in Parkinson disease. Am J Ther, 2006, 13: 445-457
27. Clapham P R, Weiss R A. Immunodeficiency viruses. Spoilt for choice of co-receptors. Nature,1997, 388: 230-231
28. Cohen E A, Subbramanian R A, Gottlinger H G. Role of auxiliary proteins in retroviral morphogenesis. Curr Top Microbiol Immunol,1996, 214: 219-235
29. Colombo S, Longhi R, Alcaro S, Ortuso F, Sprocati T, Flora A, Borgese N. N-myristoylation determines dual targeting of mammalian NADH-cytochrome b5 reductase to ER and mitochondrial outer membranes by a mechanism of kinetic partitioning. J Cell Biol, 2005,168: 735-745
30. Daoud R, Mies G, Smialowska A, Olah L, Hossmann KA, Stamm S. Ischemia induces a translocation of the splicing factor tra2-beta 1 and changes alternative splicing patterns in the brain. J Neurosci, 2002, 22: 5889-5899
31. Fire A, Xu S, Montgomery M K, Kostas S A, Driver S E, Mello C C. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 1998, 391: 806-811
32. Fleury C, Mignotte B. Vayssiere J L. Mitochondrial reactive oxygen species in cell death signaling. Biochimie, 2002, 84:131-141
33. Forrest M S, Lan Q, Hubbard A E, Zhang L, Vermeulen R, Zhao X, Li G, Wu Y Y, Shen M, Yin S, Chanock S J, Rothman N, Smith M T. Discovery of novel biomarkers by microarray analysis of peripheral blood mononuclear cell gene expression in benzene-exposed workers. Environ Health Perspect,2005,113: 801-807
34. Frankel A D, Young J A. HIV-1: fifteen proteins and an RNA. Annu Rev Biochem, 1998, 67: 1-25
35. Gerolami R, Uch R, Faivre J, Garcia S, Hardwigsen J, Cardoso J, Mathieu S, Bagnis C, Brechot C, Mannoni P. Herpes simplex virus thymidine kinase-mediated suicide gene therapy for hepatocellular carcinoma using HIV-1-derived lentiviral vectors. J Hepatol, 2004, 40: 291-297
36. Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H. Transcriptional activation by tetracyclines in mammalian cells. Science,1995, 268: 1766-1769
37. Griffith T S, Rauch C T, Smolak P J, Waugh J Y, Boiani N, Lynch D H, Smith C A, Goodwin R G, Kubin M Z. Functional analysis of TRAIL receptors using monoclonal antibodies. J Immunol, 1999,162: 2597-2605
38. Gutierrez-Adan A, Pintado B. Effect of flanking matrix attachment regions on the expression of microinjected transgenes during preimplantation development of mouse embryos. Transgenic Res, 2000, 9: 81-89
39. Hamada T, Sasaki H, Seki R, Sakaki Y. Mechanism of chromosomal integration of transgenes in microinjected mouse eggs: sequence analysis of genome-transgene and transgene-transgene junctions at two loci. Gene, 1993,128: 197-202
40. Hamra F K, Gatlin J, Chapman K M, Grellhesl D M, Garcia J V, Hammer R E, Garbers D L. Production of transgenic rats by lentiviral transduction of male germ-line stem cells. Proc Natl Acad Sci U S A,2002, 99:14931-14936
41. Hanamura A, Caceres J F, Mayeda A, Franza B R, Jr, Krainer A R. Regulated tissue-specific expression of antagonistic pre-mRNA splicing factors. Rna, 1998, 4: 430-444
42. Harrison P M, Kumar A, Lang N, Snyder M, Gerstein M. A question of size: the eukaryotic proteome and the problems in defining it. Nucleic Acids Res, 2002, 30: 1083-1090
43. Hawken R J, Murtaugh J, Flickinger G H, Yerle M, Robic A, Milan D, Gellin J, Beattie C W, Schook L B, Alexander LJ. A first-generation porcine whole-genome radiation hybrid map. Mamm Genome, 1999,10: 824-830
44. Hernandez L D, Hoffman L R, Wolfsberg T G, White J M. Virus-cell and cell-cell fusion. Annu Rev Cell Dev Biol, 1996,12: 627-661
45. Hofmann A, Kessler B, Ewerling S, Weppert M, Vogg B, Ludwig H, Stojkovic M, Boelhauve M, Brem G, Wolf E, Pfeifer A. Efficient transgenesis in farm animals by lentiviral vectors. EMBO Rep,2003, 4: 1054-1060
46. Honaramooz, A, Behboodi E, Blash S, Megee S O, Dobrinski I. Germ cell transplantation in goats. Mol Reprod Dev,2003, 64: 422-428
47. Honaramooz A, Megee S O, Dobrinski I. Germ cell transplantation in pigs. Biol Reprod, 2002, 66: 21-28
48. Indraccolo S, Habeler W, Tisato V, Stievano L, Piovan E, Tosello V, Esposito G, Wagner R, Uberla K, Chieco-Bianchi L, Amadori A. Gene transfer in ovarian cancer cells: a comparison between retroviral and lentiviral vectors. Cancer Res,2002, 62: 6099-6107
49. Indraccolo S, Tisato V, Tosello V, Habeler W, Esposito G, Moserle L, Stievano L, Persano L, Chieco-Bianchi L, Amadori A. Interferon-alpha gene therapy by lentiviral vectors contrasts ovarian cancer growth through angiogenesis inhibition. Hum Gene Ther, 2005,16: 957-970
50. Izadyar F, Den Ouden K, Stout T A, Stout J, Coret J, Lankveld D P, Spoormakers T J, Colenbrander B, Oldenbroek JK, Van der Ploeg K D, Woelders H, Kal H B, De Rooij D G. Autologous and homologous transplantation of bovine spermatogonial stem cells. Reproduction, 2003,126: 765-774
51. Jaenisch R. Germ line integration and Mendelian transmission of the exogenous Moloney leukemia virus. Proc Natl Acad Sci U S A,1976, 73:1260-1264
52. Kafri T, van Praag H, Gage F H, Verma I M. Lentiviral vectors: regulated gene expression. Mol Ther, 2000,1: 516-521
53. Kampa D, Cheng J, Kapranov P, Yamanaka M, Brubaker S, Cawley S, Drenkow J, Piccolboni A, Bekiranov S, Helt G, Tammana H, Gingeras T R. Novel RNAs identified from an in-depth analysis of the transcriptome of human chromosomes 21 and 22. Genome Res, 2004,14: 331-342
54. Kordower J H. In vivo gene delivery of glial cell line-derived neurotrophic factor for Parkinson's disease. Ann Neurol 53 Suppl 3, 2003, S120-132; discussion S132-134
55. Krawczak M, Reiss J, Cooper D N. The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: causes and consequences. Hum Genet, 1992, 90: 41-54
56. Lai Z, Brady R O. Gene transfer into the central nervous system in vivo using a recombinanat lentivirus vector. J Neurosci Res, 2002, 67: 363-371
57. Lamb R A, Pinto L H. Do Vpu and Vpr of human immunodeficiency virus type 1 and NB of influenza B virus have ion channel activities in the viral life cycles? Virology, 1997,229: 1-11
58. Lander E S, Linton L M, Birren B, Nusbaum C, Zody M C, Baldwin J, Devon K, Dewar K, Doyle M, FitzHugh W, Funke R, Gage D, Harris K, Heaford A, Howland J, Kann L, Lehoczky J, LeVine R, McEwan P, McKernan K, Meldrim J, Mesirov J P, Miranda C, Morris W, Naylor J, Raymond C, Rosetti M, Santos R, Sheridan A, Sougnez C, Stange-Thomann N, Stojanovic N, Subramanian A, Wyman D, Rogers J, Sulston J, Ainscough R, Beck S, Bentley D, Burton J, Clee C, Carter N, Coulson A, Deadman R, Deloukas P, Dunham A, Dunham I, Durbin R, French L, Grafham D, Gregory S, Hubbard T, Humphray S, Hunt A, Jones M, Lloyd C, McMurray A, Matthews L, Mercer S, Milne S, Mullikin J C, Mungall A, Plumb R, Ross M, Shownkeen R, Sims S, Waterston R H, Wilson R K, Hillier L W, McPherson J D, Marra M A, Mardis E R, Fulton L A, Chinwalla AT, Pepin K H, Gish W R, Chissoe S L, Wendl M C, Delehaunty K D, Miner T L, Delehaunty A, Kramer J B, Cook L L, Fulton RS, Johnson D L, Minx P J, Clifton S W, Hawkins T, Branscomb E, Predki P, Richardson P, Wenning S, Slezak T, Doggett N, Cheng J F, Olsen A, Lucas S, Elkin C, Uberbacher E, Frazier M, et al. Initial sequencing and analysis of the human genome. Nature, 2001, 409: 860-921
59. Le Gall S, Heard J M, Schwartz O. Analysis of Nef-induced MHC-I endocytosis. Res Virol, 1997,148: 43-47
60. Lois C, Hong E J, Pease S, Brown E J, Baltimore D. Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science, 2002, 295: 868-872
61. Mahe D, Fischer N, Decimo D, Fuchs J P. Spatiotemporal regulation of hnRNP M and 2H9 gene expression during mouse embryonic development. Biochim Biophys Acta, 2000,1492: 414-424
62. Makarova I V, Tarantul V Z, Gazarian KG. [Structural features of the integration site of foreign DNA in the transgenic mouse genome]. Mol Biol (Mosk) , 1998, 22: 1553-1561
63. Mannherz O, Mertens D, Hahn M, Lichter P. Functional screening for proapoptotic genes by reverse transfection cell array technology. Genomics, 2006, 87: 665-672
64. Merilainen O, Hakkarainen T, Wahlfors T, Pellinen R, Wahlfors J. HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct. Int J Oncol, 2005, 27: 203-208
65. Metharom P, Ellem K A, Schmidt C, Wei M Q. Lentiviral vector-mediated tyrosinase-related protein 2 gene transfer to dendritic cells for the therapy of melanoma. Hum Gene Ther, 2001,12: 2203-2213
66. Milan D, Hawken R, Cabau C, Leroux S, Genet C, Lahbib Y, Tosser G, Robic A, Hatey F, Alexander L, Beattie C, Schook L, Yerle M, Gellin J. IMpRH server: an RH mapping server available on the Web. Bioinformatics, 2000,16: 558-559
67. Miyoshi H, Blomer U, Takahashi M, Gage F H, Verma I M. Development of a self-inactivating lentivirus vector. J Virol, 1998, 72: 8150-8157
68. Modrek B, Lee C. A genomic view of alternative splicing. Nat Genet, 2002, 30: 13-19
69. Morgan A, Burgoyne R D, Barclay J W, Craig T J, Prescott G R, Ciufo L F, Evans G J. Graham M E. Regulation of exocytosis by protein kinase C. Biochem Soc Trans,2005, 33:1341-1344
70. Nagano M, Brinster C J, Orwig K E, Ryu B Y, Avarbock M R, Brinster R L. Transgenic mice produced by retroviral transduction of male germ-line stem cells. Proc Natl Acad Sci U S A, 2001, 98:13090-13095
71. Nagano M, Shinohara T, Avarbock M R, Brinster R L. Retrovirus-mediated gene delivery into male germ line stem cells. FEBS Lett, 2000, 475: 7-10
72. Nakai K, Sakamoto H. Construction of a novel database containing aberrant splicing mutations of mammalian genes. Gene, 1994,141: 171-177
73. Naldini L, Blomer U, Gage F H, Trono D, Verma I M. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci U S A, 1996, 93: 11382-11388
74. Palmiter R D, Brinster R L. Transgenic mice. Cell, 1985, 41: 343-345
75. Pan G, O'Rourke K, Chinnaiyan A M, Gentz R, Ebner R, Ni J, Dixit V M. The receptor for the cytotoxic ligand TRAIL. Science, 1997, 276: 111-113
76. Park F, Ohashi K, Kay M A. Therapeutic levels of human factor VIII and IX using HIV-1-based lentiviral vectors in mouse liver. Blood,2000, 96:1173-1176
77. Pawliuk R, Westerman K A, Fabry M E, Payen E, Tighe R, Bouhassira E E, Acharya S A, Ellis J, London I M, Eaves C J, Humphries R K, Beuzard Y, Nagel R L, Leboulch P. Correction of sickle cell disease in transgenic mouse models by gene therapy. Science, 2001, 294: 2368-2371
78. Pfeifer A, Ikawa M, Dayn Y, Verma I M. Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proc Natl Acad Sci U S A, 2002, 99: 2140-2145
79. Ponder K P. Gene therapy for hemophilia. Curr Opin Hematol, 2006,13: 301-307
80. Puglisi J D, Tan R, Calnan B J, Frankel A D, Williamson JR. Conformation of the TAR RNA-arginine complex by NMR spectroscopy. Science, 1992, 257: 76-80
81. Reiser J, Lai Z, Zhang X Y, Brady R O. Development of multigene and regulated lentivirus vectors. J Virol, 2000, 74:10589-10599
82. Ribas A. Genetically modified dendritic cells for cancer immunotherapy. Curr Gene Ther, 2005, 5: 619-628
83. Rinn J L, Euskirchen G, Bertone P, Martone R, Luscombe N M, Hartman S, Harrison P M, Nelson F K, Miller P, Gerstein M, Weissman S, Snyder M. The transcriptional activity of human Chromosome 22. Genes Dev, 2003,17: 529-540
84. Rozovskaia T, Feinstein E, Mor O, Foa R, Blechman J, Nakamura T, Croce C M, Cimino G. Canaani E. Upregulation of Meisl and HoxA9 in acute lymphocytic leukemias with the t (4 :11) abnormality. Oncogene, 2001, 20: 874-878
85. Russo I H, Russo J. Hormonal approach to breast cancer prevention. J Cell Biochem Suppl, 2000, 34:1-6
86. Scorilas A, Kyriakopoulou L, Katsaros D, Diamandis E P. Cloning of a gene (SR-A1) , encoding for a new member of the human Ser/Arg-rich family of pre-mRNA splicing factors: overexpression in aggressive ovarian cancer. Br J Cancer, 2001, 85: 190-198
87. Song Y, Zhang Z, Yu X, Yan M, Zhang X, Gu S, Stuart T, Liu C, Reiser J, Zhang Y, Chen Y. Application of lentivirus-mediated RNAi in studying gene function in mammalian tooth development. Dev Dyn, 2006, 235:1334-1344
88. Stahlberg H, Fotiadis D, Scheuring S, Remigy H, Braun T, Mitsuoka K, Fujiyoshi Y, Engel A. Two-dimensional crystals: a powerful approach to assess structure, function and dynamics of membrane proteins. FEBS Lett, 2001, 504:166-172
89. Stamm S. Signals and their transduction pathways regulating alternative splicing: a new dimension of the human genome. Hum Mol Genet, 2002,11: 2409-2416
90. Stelzl U, Worm U, Lalowski M, Haenig C, Brembeck F H, Goehler H, Stroedicke M, Zenkner M, Schoenherr A, Koeppen S, Timm J, Mintzlaff S, Abraham C, Bock N, Kietzmann S, Goedde A, Toksoz E, Droege A, Krobitsch S, Korn B, Birchmeier W, Lehrach H, Wanker E E. A human protein-protein interaction network: a resource for annotating the proteome. Cell, 2005,122: 957-968
91. Stickeler E, Kittrell F, Medina D, Berget S M. Stage-specific changes in SR splicing factors and alternative splicing in mammary tumorigenesis. Oncogene, 1999, 18: 3574-3582
92. Stutz F, Neville M, Rosbash M. Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast. Cell, 1995, 82: 495-506
93. Sun M, Zhang G R, Kong L, Holmes C, Wang X, Zhang W, Goldstein D S, Geller AI. Correction of a rat model of Parkinson's disease by coexpression of tyrosine hydroxylase and aromatic amino acid decarboxylase from a helper virus-free herpes simplex virus type 1 vector. Hum Gene Ther, 2003,14: 415-424
94. Ullman K S, Powers M A, Forbes D J. Nuclear export receptors: from importin to exportin. Cell, 1997, 90: 967-970
95. Vaux D L. Caspases and apoptosis - biology and terminology. Cell Death Differ, 1999, 6: 493-494
96. Venter J C, Adams M D, Myers E W, Li P W, Mural R J, Sutton G G, Smith H O, Yandell M, Evans C A, Holt R A, Gocayne J D, Amanatides P, Ballew R M, Huson D H, Wortman J R, Zhang Q, Kodira C D, Zheng X H, Chen L, Skupski M, Subramanian G, et al. The sequence of the human genome. Science, 2001, 291: 1304-1351
97. Walczak H, Degli-Esposti M A, Johnson R S, Smolak P J, Waugh J Y, Boiani N, Timour M S, Gerhart M J, Schooley K A, Smith C A, Goodwin R G, Rauch C T. TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL. Embo J, 1997, 16: 5386-5397
98. Yamane K, Kinsella T J. Casein kinase 2 regulates both apoptosis and the cell cycle following DNA damage induced by 6-thioguanine. Clin Cancer Res, 2005, 11: 2355-2363
99. Yerle M, Pinton P, Robic A, Alfonso A, Palvadeau Y, Delcros C, Hawken R, Alexander L, Beattie C, Schook L, Milan D, Gellin J. Construction of a whole-genome radiation hybrid panel for high-resolution gene mapping in pigs. Cytogenet Cell Genet, 1998, 82:182-188
100.Yu X, Zhan X, D'Costa J, Tanavde V M, Ye Z, Peng T, Malehorn M T, Yang X, Civin C I Cheng L. Lentiviral vectors with two independent internal promoters transfer high-level expression of multiple transgenes to human hematopoietic stem-progenitor cells. Mol Ther, 2003, 7: 827-838
101.Zheng J, Kitajima K, Sakai E, Kimura T, Minegishi N, Yamamoto M, Nakano T. Differential effects of GATA-1 on proliferation and differentiation of erythroid lineage cells. Blood, 2006,107: 520-527
102.Zoraqi G, Spadafora C. Integration of foreign DNA sequences into mouse sperm genome. DNA Cell Biol, 1997,16: 291-300