超声波辅助细胞穿透多肽介导的花粉遗传转化技术的研究
|
摘要
自世界上首例转基因烟草问世以来,植物转基因技术得到广泛应用,特别是在作物育种方面。尽管在过去的几年中玉米的转化技术取得了较大进展,但对研究者来说遗传转化技术仍然是玉米转基因研究的一个关键限制因素。目前常用的方法是基因枪法和农杆菌介导法。因此需开发一种新的遗传转化方法,建立稳定、便捷和高效的遗传转化,加速作物新品种培育的步伐和功能基因组学研究的发展。应用细胞穿透多肽(cell penetrate peptides,CPPs)为介导对花粉进行的转基因研究,无需大量准备工作,从收集花粉到完成遗传转化可在几小时内完成。本文就超声处理参数、植物表达载体构建、转基因植株草甘膦筛选浓度及转基因检测等方面做了相关研究。
主要研究结果如下:
1.以玉米自交系昌7-2为材料,利用TTC染色法和花粉离体萌发的方法研究了不同培养基组分和不同超声参数对花粉活力的影响。结果表明:花粉在液体中的活力急剧下降;随着超声处理强度的加大,花粉的活力明显降低;相同超声强度下,超声处理时间越长,花粉的活力越低。SAS软件分析表明在四种不同的培养基中,PLM1和PLM3对花粉活力的影响相对小一些。不同超声处理对花粉活力的影响:N1<1<3<4<2<5<6。为以花粉作为受体进行转基因的研究提供超声参数参考。
2.利用In-FusionTM Advantage PCR Cloning Kit(Clontech Laboratories, Takara)试剂盒,将玉米谷氨酰胺合成酶基因(GS1)插入pCUbi1390载体的Ubi启动子后面PstI酶切位点处,然后在此基础之上,再将选择标记基因5-烯醇丙酮莽草酸-3-磷酸合成酶基因( CaMV35S + EPSPS )插入HindIII位点,构建植物表达载体pCUbi1390-Ubi-GS1-35S-EPSPS。结果表明In-Fusion法正确构建了设计的转化载体,简化了载体构建的过程,也适用于任何一个基于单酶切位点把外源DNA重组插入目标序列,具有应用与推广价值。
3.以玉米自交系18-599(红)为材料,当幼苗4-5叶时,分别喷施不同浓度的草甘膦溶液,确定转基因植株草甘膦筛选浓度。幼苗在喷施0.2%的草甘膦一周后死亡77株,受抑制163株,正常9株,96.4%的非转基因玉米苗对0.2%的草甘膦没有抗性。因此,选用0.2%的草甘膦药液作为筛选转基因植株的最适浓度。
4.以玉美头168为材料,5%的蔗糖溶液为超声处理缓冲液进行花粉介导的遗传转化实验。收获种子后,对T0代进行PCR检测、Elisa检测试验。利用不同超声波处理参数处理花粉,授粉后总共得到种子467粒,将种子种到温室里共出苗432株,经过三轮的PCR检测,共有23株幼苗为转基因植株。PCR检测转基因效率约为5.32%。对可以正常结实的18株PCR检测阳性的转基因植株进行Elisa检测,18个单株全部呈现阳性。进一步转基因检测得到的转基因效率为4.17%。
Transgenic technology has been widely used since its inception of the first transgenic plant--transgenic tobacco, especially in the area of crop breeding. Although the maize genetic transformation has been made great progresses in the recent years, it still the vital limitation. Particle bombardment and agrobacterium-mediated techniques are commonly used these days. It’s necessary to develop new transformation methods and built stable, rapid and high-efficient transformation system for breeding new crop variety and the study of functional genomics. With the help of cell penetrating peptides, pollen-mediated transformation need not lots of preparatory work. And the complete genetic transformation can be completed in a few hours. In this paper, ultrasonic processing parameters, construction of plant expression vector, transgenic plants glyphosate screening concentration and transgenic detection were studied.
The results were as follows:
1. We used the maize inbred line Chang 7-2 to study the viability of maize pollen in different media components and different ultrasonic parameters with the methods TTC staining and pollen germination in vitro. The results showed that: The viability of pollen was sharply decreased in liquid. With the increasing intensity of ultrasound treatment, the viability of pollen gradually decreased; and also the time of ultrasound treatment. Analysis of SAS confirmed that PLM1 and PLM3 had less effect on pollen viability among the four media; and the influence of different ultrasound treatment was: N1<1<3<4<5<6<2. This work can provide ultrasound parameters reference for pollen-mediated transgenic research.
2. We inserted two genes: maize glutamine synthase gene (GS1) and 5-enolpyruvlshimimate-3-phosphate synthase gene (CaMV35S + EPSPS ) into the Pst I and Hind III sites of vector pCUbi1390 separately to construct plant expression vector pCUbi1390-Ubi-GS1-35S-EPSPS. The results showed the expecting pCUbi1390-Ubi-GS1-35S-EPSPS was successfully constructed. The procedures were largely simplified and the method could be widely used in any recombination of insertion of any DNA sequence not very long at single restriction site of any DNA sequence.
3. Used maize inbred line 18-599(red) as the material, we sprayed 4-5 leaf seedling glyphosate solution of different concentrations to determine the transgenic plants, glyphosate screening concentration. Seedlings of spraying 0.2% glyphosate had 77 death, 163 inhibition and 9 normal. That said 96.4% of seedling had no resistance to 0.2% glyphosate solution. So we choose 0.2% as the optimal concentration to screen Transgenic plants.
4. The pollen-mediated genetic transformation was carried out using Yumeitou 168 as material, 5% sucrose solution for ultrasound processing buffer. After harvest, we did PCR amplification analysis and Elisa test on T0 generation seedlings to detect the transgenic efficiency of this method. Sowed the seeds we harvested (467 in total), 432 seeds emerged. 23 seedlings were positive after PCR of three pair of primers, and the transgenic efficiency were approximately 5.32%. 18 of PCR positive plants which were fertile were assayed again with Elisa test. They all tested positive. Further transgenic detected of transgenic efficiency was 4.17%.
主要研究结果如下:
1.以玉米自交系昌7-2为材料,利用TTC染色法和花粉离体萌发的方法研究了不同培养基组分和不同超声参数对花粉活力的影响。结果表明:花粉在液体中的活力急剧下降;随着超声处理强度的加大,花粉的活力明显降低;相同超声强度下,超声处理时间越长,花粉的活力越低。SAS软件分析表明在四种不同的培养基中,PLM1和PLM3对花粉活力的影响相对小一些。不同超声处理对花粉活力的影响:N1<1<3<4<2<5<6。为以花粉作为受体进行转基因的研究提供超声参数参考。
2.利用In-FusionTM Advantage PCR Cloning Kit(Clontech Laboratories, Takara)试剂盒,将玉米谷氨酰胺合成酶基因(GS1)插入pCUbi1390载体的Ubi启动子后面PstI酶切位点处,然后在此基础之上,再将选择标记基因5-烯醇丙酮莽草酸-3-磷酸合成酶基因( CaMV35S + EPSPS )插入HindIII位点,构建植物表达载体pCUbi1390-Ubi-GS1-35S-EPSPS。结果表明In-Fusion法正确构建了设计的转化载体,简化了载体构建的过程,也适用于任何一个基于单酶切位点把外源DNA重组插入目标序列,具有应用与推广价值。
3.以玉米自交系18-599(红)为材料,当幼苗4-5叶时,分别喷施不同浓度的草甘膦溶液,确定转基因植株草甘膦筛选浓度。幼苗在喷施0.2%的草甘膦一周后死亡77株,受抑制163株,正常9株,96.4%的非转基因玉米苗对0.2%的草甘膦没有抗性。因此,选用0.2%的草甘膦药液作为筛选转基因植株的最适浓度。
4.以玉美头168为材料,5%的蔗糖溶液为超声处理缓冲液进行花粉介导的遗传转化实验。收获种子后,对T0代进行PCR检测、Elisa检测试验。利用不同超声波处理参数处理花粉,授粉后总共得到种子467粒,将种子种到温室里共出苗432株,经过三轮的PCR检测,共有23株幼苗为转基因植株。PCR检测转基因效率约为5.32%。对可以正常结实的18株PCR检测阳性的转基因植株进行Elisa检测,18个单株全部呈现阳性。进一步转基因检测得到的转基因效率为4.17%。
Transgenic technology has been widely used since its inception of the first transgenic plant--transgenic tobacco, especially in the area of crop breeding. Although the maize genetic transformation has been made great progresses in the recent years, it still the vital limitation. Particle bombardment and agrobacterium-mediated techniques are commonly used these days. It’s necessary to develop new transformation methods and built stable, rapid and high-efficient transformation system for breeding new crop variety and the study of functional genomics. With the help of cell penetrating peptides, pollen-mediated transformation need not lots of preparatory work. And the complete genetic transformation can be completed in a few hours. In this paper, ultrasonic processing parameters, construction of plant expression vector, transgenic plants glyphosate screening concentration and transgenic detection were studied.
The results were as follows:
1. We used the maize inbred line Chang 7-2 to study the viability of maize pollen in different media components and different ultrasonic parameters with the methods TTC staining and pollen germination in vitro. The results showed that: The viability of pollen was sharply decreased in liquid. With the increasing intensity of ultrasound treatment, the viability of pollen gradually decreased; and also the time of ultrasound treatment. Analysis of SAS confirmed that PLM1 and PLM3 had less effect on pollen viability among the four media; and the influence of different ultrasound treatment was: N1<1<3<4<5<6<2. This work can provide ultrasound parameters reference for pollen-mediated transgenic research.
2. We inserted two genes: maize glutamine synthase gene (GS1) and 5-enolpyruvlshimimate-3-phosphate synthase gene (CaMV35S + EPSPS ) into the Pst I and Hind III sites of vector pCUbi1390 separately to construct plant expression vector pCUbi1390-Ubi-GS1-35S-EPSPS. The results showed the expecting pCUbi1390-Ubi-GS1-35S-EPSPS was successfully constructed. The procedures were largely simplified and the method could be widely used in any recombination of insertion of any DNA sequence not very long at single restriction site of any DNA sequence.
3. Used maize inbred line 18-599(red) as the material, we sprayed 4-5 leaf seedling glyphosate solution of different concentrations to determine the transgenic plants, glyphosate screening concentration. Seedlings of spraying 0.2% glyphosate had 77 death, 163 inhibition and 9 normal. That said 96.4% of seedling had no resistance to 0.2% glyphosate solution. So we choose 0.2% as the optimal concentration to screen Transgenic plants.
4. The pollen-mediated genetic transformation was carried out using Yumeitou 168 as material, 5% sucrose solution for ultrasound processing buffer. After harvest, we did PCR amplification analysis and Elisa test on T0 generation seedlings to detect the transgenic efficiency of this method. Sowed the seeds we harvested (467 in total), 432 seeds emerged. 23 seedlings were positive after PCR of three pair of primers, and the transgenic efficiency were approximately 5.32%. 18 of PCR positive plants which were fertile were assayed again with Elisa test. They all tested positive. Further transgenic detected of transgenic efficiency was 4.17%.
引文
陈梁鸿,王新望,张文俊,张晓东,胡道芬,刘文田. 1999.抗除草剂草甘膦EPSPs基因在小麦中的转化.遗传学报, 26(3): 239~243
陈其军,安瑞,周海梦,陈珈,王学臣. 2004.使用Gateway技术兼容的T载体获得入门克隆.生物化学与生物物理进展, 31(10): 951~954
陈茜,刘亚伟,黄邵,杨勤,姜勇. 2009.细胞穿透肽穿膜机制的研究进展.国际病理科学与临床杂志, 29(2): 115~120
邓丽琴,祝朋芳,陈长青. 2004.试论常规育种与分子育种的研究应用.杂粮作物, 24(005): 280~281
丁志山,沃兴德. 1997.超声波的生物学效用及其在转基因中的应用.生命科学, 9(4): 187~189
董云洲,段胜军,赵连元,杨秀海,贾士荣. 1999.用基因枪转化花粉获得转基因谷子和玉米.中国农业科学, 32(2): 9~13
杜春芳,刘惠民,李朋波,孙毅,李润植. 2006.花粉介导法获得油菜转基因植株研究.作物学报, 32(5): 749~754
高东尧,夏兰芹,徐兆师,马有志,李彦舫,原亚萍,李连城,陈明,孙金海. 2009.小麦WVp-1基因表达载体的Gateway技术构建及其遗传转化.麦类作物学报, 29(2): 189~194
赫福霞,郎志宏,陆伟,林敏,张杰,黄大昉. 2008.以耐草甘膦2mG2-epsps基因为选择标记的玉米转化体系的建立.生物技术通报, 5: 92~97.
何迎春,任春梅,高必达. 2001.耐草甘膦基因克隆和作物转化研究新进展.生物技术, 11(4): 38~40
胡金艳,张书祥,王晓武,武剑. 2007.基于Gateway技术的拟南芥雄性不育GHF基因表达载体的构建. 农业生物技术科学, 23(12): 45~48
梁雪莲,郭平毅,孙毅,刘惠民,王景雪,刘少翔,赵志国,唐思静. 2005.玉米3种非组培转基因方法转化外源bar基因研究.作物学报, 31(12): 1648~1653
梁雪莲,郑奕雄,庄东红,黄德强. 2006.花生CpTI基因转化----花粉介导法.花生学报, 35(4): 1~5
林朝贵,范林,陈良龙. 2010.利用In-Fusion技术构建存活素-增强型绿色荧光蛋白融合基因重组慢病毒表达载体.心血管康复医学杂志, 19(1): 10~14
林春晶,韦正乙,蔡勤安,侯敬尧,邢少辰. 2008.几种植物转基因表达载体的构建方法.生物技术, 18 (5): 84~87
刘纪麟. 2001.玉米育种学,北京:中国农业出版社
刘锡娟,刘昱辉,王志兴,王旭静,张永强. 2007.转5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)基因抗草甘膦烟草和棉花的获得.农业生物技术学报, 15(6): 958~963
楼秀芹,周骋. 2008.特洛伊肽运载核酸的研究进展.医学研究杂志, 37(2): 92~95
陆云华,马立新,蒋思婧. 2006.一种通用高效的复杂载体构建的新方法.遗传, 28(2): 212~218
欧阳平,李玉,严红,马立新. 2007.一种载体构建的新方法:一步克隆法.湖北大学学报(自然科学版), 29(2): 178~181
石清琢,姜敏. 2005.玉米转基因育种技术概述.杂粮作物, 25(4): 230~231
苏少泉. 2009.转基因抗草甘膦作物的新进展.现代农药, 8(6): 1~4
孙敬儒,王保义. 2001.强电场脉冲致细胞膜电穿孔的观察.四川大学学报:自然科学版, 38(5):748~751
杨艳芹,谢菊芳,夏利霞,侯义峰,肖磊. 2006.脉冲电场致细胞膜电穿孔的机理分析.湖北大学学报:自然科学版, 28(2): 165~167
万丙良,张献龙. 2000.基因枪介导转化水稻花粉获得转基因植株.农业生物技术学报, 8(4): 332, 381
王宏伟,梁业红,史振声,张世煌. 2007.作物抗草甘膦转基因研究概况术.作物杂志, 4: 9~12
王节之,郝晓芬,郑向阳,王路英,孙毅,王景雪. 2004.谷子花粉介导法转几丁质酶基因的研究.生物技术, 14(5): 5~6
王景雪,孙毅,崔贵梅,胡晶晶. 2001.花粉介导法获得玉米转基因植株.植物学报, 43(3): 275~279
王秀君,郎志宏,陆伟,林敏,单安山,黄大昉. 2008.利用花粉管通道法将耐草甘膦基因mG2-epsps导入玉米自交系的研究初报.中国农业科技导报, 10(4): 56~62
徐化学,熊建华,傅彬英. 2007.应用Gateway技术构建水稻OsDAD1基因的RNA干扰载体.分子植物育种, 5(1): 133~136
张宏,王国英,谢友菊,戴景瑞,许宁,赵南明,李太源,田颖川,乔利亚,莽克强. 1997.超声波介导
转化玉米愈伤组织及可育转基因植株的获得.中国科学(C辑), 27(2): 162~167
张素芝,荣廷昭. 2008.农杆菌介导的玉米遗传转化系统研究进展.遗传, 30(10): 1249~1256
郑秀芳,金治平. 2002.植物遗传转化技术研究概况.河西学院学报, 5: 66~70
周双林,夏苗芬. 2006.肽核酸的研究与应用.中国药学杂志, 41(15): 1124~1128
朱玉. 2002.极端污染环境下草甘膦耐受菌株荧光假单胞菌G2的鉴定及其EPSP合酶基因的克隆. [硕士学位论文].北京:中国农业科学院
朱玉,于中连,林敏. 2003.草甘膦生物抗性和生物降解及其转基因研究.分子植物育种, 1(4): 435~441
Albarran B, To R, Stayton P S. 2005. A TAT-streptavidin fusion protein directs uptake of biotinylated cargo into mammalian cells. Protein Engineering Design and Selection, 18(3): 147~152
Arnizen C J, DeBonte Jr L R, Evans D A, Loh W H, O'dell J T. 1991. 1991-09-17. Pollen-mediated gene transformation in plants. United States Patents, 5049500
Astriab-Fisher A, Sergueev D, Fisher M, Shaw B R, Juliano R L. 2002. Conjugates of antisense oligonucleotides with the Tat and Antennapedia cell-penetrating peptides: effects on cellular uptake, binding to target sequences and biologic actions. Pharmaceutical Research, 19(6): 744~754
Benoit R M, Wilhelm R N, Scherer-Becker D, Ostermeier C. 2006. An improved method for fast, robust, and seamless integration of DNA fragments into multiple plasmids. Protein Expression and Purification, 45(1): 66~71
Berrow N S, Alderton D, Owens R J. 2009. The precise engineering of expression vectors using high-throughput In-Fusion PCR cloning. Methods in Molecular Biology, 498: 75~90.
Berrow N S, Alderton D, Sainsbury S, Nettleship J, Assenberg R, Rahman N, Stuart D I, Owens R J. 2007. A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Research, 35(6): e45
Booy G, Krens F A, Huizing H J. 1989. Attempted pollen-mediated transformation of maize. Journal of Plant Physiology, 135(3): 319~324
Brewbaker J L, Kwack B H. 1963. The essential role of calcium ion in pollen germination and pollen tube growth. American Journal of Botany, 50(9): 859~865
Brooks H, Lebleu B, Vives E. 2005. Tat peptide-mediated cellular delivery: back to basics. Advanced DrugDelivery Reviews, 57(4): 559~577
Chang M, Chou J C, Lee H J. 2005. Cellular internalization of fluorescent proteins via arginine-rich intracellular delivery peptide in plant cells. Plant and Cell Physiology, 46(3): 482~488
Chiu Y L, Ali A, Chu C Y, Cao H, Rana T M. 2004. Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells. Chemistry & Biology, 11: 1165~1175
Chowrira G M, Akella V, Lurquin P F. 1995. Eletroporation-mediated gene transfer into intact nodal meristems in planta: generating transgenic plants without in vitro tissue culture. Molecular Biotechnology, 3(1): 17~23
Chugh A, Amundsen E, Eudes F. 2009. Translocation of cell-penetrating peptides and delivery of their cargoes in triticale microspores. Plant Cell Reports, 28(5): 801~810
Chugh A, Eudes F. 2007. Translocation and nuclear accumulation of monomer and dimer of HIV-1 Tat basic domain in triticale mesophyll protoplasts. Biochimica et Biophysica Acta, 1768(3): 419~426
Chugh A, Eudes F. 2008. Study of uptake of cell penetrating peptides and their cargoes in permeabilized wheat immature embryos. FEBS Journal, 275(10):2403~2414
Curtis M D, Grossniklaus V. 2003. A gateway cloning vector site for high-throughput functional analysis of genes in planta. Plant Physiology, 133(2): 462~469
D’Hallui K, Bonne E, Bossut M, De Beuckeleer M, Leemans J. 1992. Transgenic maize plants by tissue electroporation. Plant Cel1, 4(12): 1495~1505
Dietz G P H, Bahr M. 2007. Synthesis of cell-penetrating peptides and their application in neurobiology. Neuroprotection Methods and Protocols, 399: 181~198
Dill G M. 2005. Glyphosate-resistant crops: history, status and future. Pest Management Science, 61(3): 219~224
Dong J, Kharb P, Teng W, Hall T C. 2001. Characterization of rice transformed via an Agrobacterium-mediated inflorescence approach. Molecular Breeding, 7(3): 187~194
Duke S O. 2005. Taking stock of herbicide-resistant crops ten years after introduction. Pest Management Science, 61(3): 211~218
Fawell S, Seery J, Daikh Y, Moore C, Chen L L, Pepinsky B, Barsoum J. 1994. Tat-mediated delivery of heterologous proteins into cells. Proceedings of the National Academy of Sciences of the United States of America, 91(2): 664~668
Ferrari A, Pellegrini V, Arcangeli C, Fittipaldi A, Giacca M, Beltram F. 2003. Caveolae-mediated internalization of extracellularHIV-1 Tat fusion proteins visualized in real time. Molecular Therapy, 899(2): 284~293
Fittipaldi A, Ferrari A, Zooppe M, Arcangeli C, Pellegrini V, Beltram F, Giacca M. 2003. Cell membrane lipid rafts mediated caveolar endocytosis of HIV-1 Tat fusion proteins. Journal of Biological
Chemistry, 278(36): 34141~34149 Fominaya J, Gasset M, Garcia R, Roncal F, Albar J P, Bernad A. 2000. An optimised amphiphilic cationic peptide as an efficient non-viral gene delivery vector. Journal of Gene Medicine, 2(6): 455~464
Frankel A D, Pabo C O. 1988. Cellular uptake of the tat protein from human immunodeficiency virus. Cell, 55(6): 1189~1193
Fukuoka H, Ogawa T, Mitsuhara I, Iwai T, Isuzigawa K, Nishizawa Y, Gotoh Y, Nishizawa Y, Tagiri A, Ugaki M, Ohshiama M, Yano H, Murai N, Niwa Y, Hibi T, Ohashi Y. 2000. Agrobacterium-mediatedtransformation of monocot and dicot plants using the NCR promoter derived from soybean chlorotic mottle virus. Plant Cell Reports, 19(8): 815~820
Futaki S. 2006. Oligoarginine vectors for intracellular delivery: Design and cellular?\uptake mechanisms. Peptide Science, 84(3): 241~249
Grimsley N, Hohn T, Davies J W, Hohn B. 1987. Agrobacterium-mediated delivery of infectious maize streak virus into maize plants. Nature, 325(8): 177~179
Gordon-Kamm W J, Spencer T M, Mangano M L, Adams T R, Daines R J, Start W G, O’Brien J V, Chambers S A, Adams Jr W R, Willetts N C, Rice T B, Mackey C J, Krueger R W, Kausch A P, Lemaux PG. 1990. Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell, 2(7): 603~618
Gould J, Devey M, Hasegawa O, Ulian E C, Peterson G, Smith R H. 1991. Transformation of Zea mays L. using Agrobacterium tumefaciens and the shoot apex. Plant Physiology, 95(2): 426~434
Gasser C S, Fraley R T. 1989. Genetically Engineering Plants for Crop Improvement. Science, 244(4910): 1293~1299
Green M, Loewenstein P M. 1988. Autonomous functional domains of chemically synthesized human immunodeficiency virus tat trans-activator protein. Cell, 55(6): 1179~1188
Green J M. 2007. Review of glyphosate and ALS-inhibiting herbicide crop resistance and resistant weed management. Weed Technology, 21(2): 547~558
Hallui K, Bonne E, Bossut M, Beuckeleer M D, Leemans J. 1992. Transgenic maize plants by tissue electroporation. Plant Cell, 4(12): 1495~1505
Hamilton D A, Roy M, Rueda J , Sindhu R K, Sanford, Mascarenhas J P. 1992. Dissection of a pollen-specific promoter from maize by transient transformation assays. Plant Molecular Biology, 18(2): 211~218
Hansen G, Wright M S. 1999. Recent advances in the transformation of plants. Trends in Plant Science, 4(6): 226~231
Hartley J L, Temple G F, Brasch M A. 2000. DNA cloning using in vitro site-specific recombination. Genome Research, 10: 1788~1795
Hay L, Lachance D, Vonaderkas P, Charest P J. 1994. Transient chimeric gene expression in pollen of five conifer species following micmprojectile bombardment. Canadian Journal of Forest Research, 24: 2417~2423
Hellgren I, Gorman J, Sylven C. 2004. Factors controlling the efficiency of Tat-mediated plasmid DNA transfer. the Journal of Drug Targeting, 12(1): 39~47
Hess, D., Gresshoff, P.M., Fielitz, U., Gleiss, D. 1974. Uptake of protein and bacteriophage into swelling and germinating pollen of Petunia hybrida. Z. Pflanzenphysiol, 74: 371~376
Hess D, Lorz H. 1974. Uptake of bacterial DNA into swelling and germinating pollen grains of Petunia hybrida and Nicotiana glauca. Z. Pflanzenphysiol, 74: 52~63
Hess D. 1977. Cell modification by DNA uptake. In: Reinert J, Bajaj YPS (eds) Applied and fundamental aspects of plant cell, tissue, and organ culture. Springer-Verlag, Berlin Heidelberg, pp506~577
Hess D. 1980. Investigations on the intra- and interspecific transfer of anthocyanin genes using pollen as vectors. Z Pflanzenphysiol, 98: 321~337
Hess D. 1987. Pollen-based techniques in genetic manipulation. International Reciew of Cytology, 107:367~395
Ishida Y, Saito H, Ohta S, Hiei Y, Komari T, Kumashiro T. l996. High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens. Nature Biotechnology, l4(6): 745~750
Jacob G S, Garbow O R, Hallas L E, Kimack N M, Kishore G M Schaefer J.1988. Metabolism of Glyphosate in Pseudomonas sp. Strain LBr. Applied and Environmental Microbio1ogy, 54(12):
2953~2958 Jeang K T, Xiao H, Rich E A. 1999. Multifaceted activities of the HIV-1 transactivator of transcription, Tat. the Journal of Biological Chemistry, 274(41): 28837~28840
Joersbo M, Brunstedt J. 1990. Direct gene transfer to plant protoplasts by mild sonication. Plant Cell Reports, 9(4): 207~210
Karimi M, Depicker A and Hilson P. 2007. Recombinational cloning with plant gateway vectors. Plant Physiology, 145(4): 1144~1154
Klein T M, Fromm M, Weissinger A, Tomes D, Schaaf S, Sletten M, Sanford J C. 1988. Transfer of foreign genes into intact maize cells with high-velocity microprojectiles. Proceedings of the National Academy of Sciences of the United States of America, 85(12): 4305~4309
Koppelhus U, Awasthi S K, ZacharV, Holst H U, Ebbesen P, Nielsen P E. 2002. Cell-dependent differential cellularuptake of PNA, peptides and PNA-peptide conjugates. Antisense Nucleic Acid Drug Development, 12(2): 51~63
Landy A. 1989. Dynamic, structural and regulatory aspects ofλsite-specific recombination. Annual Reviews of Biochemistry, 58: 913~941
Lansac A R, Sullivan C Y, Johnson B E, Lee K W. 1994. Viability and germination of the pollen of sorghum [Sorghum bicolor (L.) Moench]. Annals of Botany, 74(1): 27~33
Li Y H, Tremblay F M, Séguin A. 1994. Transient transformation of pollen and embryogenic tissues of white spruce (Picea glauca(Moench.) Voss) resulting from microprojectile bombardment. Plant Cell Reports, 13(12): 66l~665
Lindsay M A. 2002. Peptide-mediated cell delivery: application in protein target validation. Current Opinion in Pharmacology, 2(5): 587~594
Loret E P, Vives E, Ho P S, Rochat H, Van Rietschoten J, Johnson Jr W C. 1991. Activating region of HIV-1 Tat protein: vacuum UV circular dichroism and energy minimization. Biochemistry, 30(24): 6013~6023
Marsischky G, LaBaer J. 2004. Many paths to many clones: a comparative look at high-throughput cloning methods. Genome Research, 14: 2020~2028.
Martinussen I, Junttila O, Twell D. 1994. Optimization of transient gene expression in pollen of Norway spruce (Picea abies) by particle acceleration. Physiologia Plantarum, 92(3): 412~416
Matthews B F, Abdul-Baki A A, Saunders J A. 1990. Expression of a foreign gene in electroporated pollen grains of tobacco. Sexual Plant Reproduction, 3(3): 147~151
McCormick S, Yamaguchi J, Twell D. 1991. Deletion analysis of pollen-expressed promoters. In Vitro Cellular & Developmental Biology-Plant, 27(1): 15~20
McCabe D, Christou P. 1993. Direct DNA transfer using electric discharge particle acceleration (ACCELLTM technology). Plant Cell, Tissue and Organ Culture, 33(3): 227~236
Nafziger E D, Widholm J M, Steinrucken H C, Killmer J L. 1984. Selection and characterization of a carrotcell line tolerant to glyphosate. Plant Physiology, 76(3): 571~574
Nishihara M, Ito M, Tanaka l, Kyo M, Ono K, Irifune K, Morikawa H. 1993. Expression of theβ-glucuronidase gene in pollen of lily(Lilium longiflorum),tobacco (Nicotiana tabacum),Nicotiana rustica,and peony (Paeonia lactiflora)by paricle Bombardment. Plant Physiology, 102: 357~361
Ohta Y. 1986. High-efficiency genetic transformation of maize by a mixture of pollen and exogenons DNA. Proceedings of the National Academy of Sciences of the United States of America, 83: 715~719
Penaloza A, Mena G L, Herrera L, Bailey A M. 1995. Cloning and sequencing of the genes involved in glyphosate utilization by Pseudomonas pseudomallei. Applied Environmental Microbiology, 61(2): 538~543
Potrykus I. 1990. Gene transfer to plants: assessment and perspectives. Physiologia Plantarum, 79(1): 125~134
Rhodes C A, Pierce D A, Mettler I J, Mascarenhas D, Detmer J J. 1988. Genetically transformed maize plants form protoplasts. Science, 240(4849): 204~207
Ruben S, Perkins A, Purcell R, Joung K, Sia R, Burghoff R, Haseltine W, Rosen C. 1989. Structural and functional characterization of human immunodeficiency virus tat protein. Journal of Virology, 63(1): 1~8
Rudolph C, Planks C, Lausier J, Schillinger U, Muller R H, Rosenecker J. 2003. Oligomers of the arginine-rich motif of the HIV-1TAT protein are capable of transferring plasmid DNA into cells. the Journal of Biological Chemistry. 278(13): 11411~11418
Saalik P, Elmquist A, Hansen M, Padari K, Saar K, Viht K, Langel U, Pooga M. 2004. Protein cargo delivery properties of cell-penetrating peptides. A comparative study. Bioconjugate Chemistry, 15(6): 1246~53
Sanford J C, Skubik K. 1986. Attempted pollen-mediated transformation using Ti-plasmids. Springer-Verlag, New York (USA), US8719725
Sanford J C, Klein T M, Wolf E D, Allen N. 1987. Delivery of substances into cells and tissues using a particle bombardment process. Particulate Science and technology, 5(1): 27~37
Sanford J C , Skubik K A , Reishch B I. 1985. A ttempted pollen-mediated plant transformation employing genomic donor DNA. Theoretical and Applied Genetics, 69(5-6): 571~574
Sasaki Y, Sane T, Yoshida S, Yahata K, Hotta T, Chesnut J D, Honda T, Imamoto F. 2004. Evidence for high specificity and efficiency of multiple recombination signals in mixed DNA cloning by the Multisite Gateway system. Journal of Biotechnology, 107(3): 233~243
Schwarze S R, Dowdy S F. 2000. In vivo protein transduction: intracellular delivery of biologically active proteins, compounds and DNA. Trends in Pharmacological Sciences, 21(2): 45~48
Sidorov V, Gilbertson L, Addae P, Duncan D. 2006. Agrobacterium-mediated transformation of seedling-derived maize callus. Plant Cell Reports, 25(4): 320~332
Sleight S C, Bartley B A, Lieviant J A, Sauro H M. 2010. In-Fusion BioBrick assembly and re-engineering. Nucleic Acids Research, 38 (8): 2624~2636
Smith C R, Saunders J A, Wer S V, Cheng J P, Matthews B F. 1994. Expression of GUS and CAT activities using electrotransformed pollen. Plant Science, 104(1): 49~58
Stoger E, Moreno R M B, Ylstra B, Vicente O, Heberle-Bors E. 1992. Comparison of different techniques for gene transfer into mature and immature tobacco pollen. Transgenic Research, 1(2): 71~78
Tan S, Evans R, Singh B. 2006. Herbicidal inhibitors of amino acid biosynthesis and herbicide-tolerant crops. Amino Acids, 30(2): 195~204
Tenno T, Goda N, Tateishi Y, Tochio1 H, Mishima M, Hayashi H, Shirakawa M, Hiroaki H. 2004. High-throughput construction method for expression vector of peptides for NMR study suited for isotopic labeling. Protein Engineering, Design & Selection, 17(4): 305~314
Torchilin V P, Levchenko T S, Rammohan R, Volodina N, Papahadjopoulos-Sternberg B, D’Souza G G M. 2003. Cell transfection in vitro and in vivo with non-toxic TAT peptide-liposome-DNA complexes. Proceedings of the National Academay of Sciences of the United States of the America, 100(4): 1972~1977
Torchilin V P, Rammohan R, Weissig V, Levchenko T S. 2001. Tat peptide on the surface of liposomes sffords their efficient intracellular delivery even at low temperature and in the presence of metabolic inhibitors. Proceedings of the National Academy of Sciences of the United States of America, 98(15): 8786~8791
Tung C H, Mueller S, Weissleder R. 2002. Novel branching membrane translocational peptide as gene delivery vector. Bioorganic & Medicinal Chemistry, 10(11): 3609~3614
Turner R , Bate N, Twell D, Foster G D. 1994. In vivo characterisation of a translational enhancer upstream from the coat protein open reading frame of potato virus S. Archives of Virology, 137: 123~132
Twell D, Klein T M., Fromm M E., McCormick S. 1989. Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiology, 91(4): 1270~1274
Twell D, Patel S, Sorensen A, Roberts M, Scott R, Draper J, Foster G. 1993. Activation and developmental regulation of an Arabidopsis anther-specific promoter in micmpores and pollen of Nicotiana tabacum. Sexual Plant Reproduction, 6: 217~224
Unnamalai N, Kang B G, Lee W S. 2004. Cationic oligopeptide-mediated delivery of dsRNA for post-transcriptional gene silencing in plant cells. FEBS Letters, 566(1-3): 307~319
van der Leede-Plegt L M, van de Ven B C E, Bino R J, van der Salm T P M, van Tunen A. 1992. Introduction and differential use of various promoters in pollen grains of Nicotiana glutinosa and Lilium longiflorum. Plant Cell Reports, 11(1): 20~24
van der Leede-Plegt L M , van de Ven B C E , Schilder M, Franken J, van Tunen A J. 1995. Development of a pollen-mediated transformation method for Nicotiana glutinosa. Transgenic Research, 4(2): 77~86
Vivès E, Brodin P and Lebleu B. 1997. A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. Journal of Biological Chemistry, 272(25): 16010~16017
Wadia J S, Dowdy S F. 2002. Protein transduction technology. Current opinion in biotechnology, 13(1): 52~56
Wang J X, Li Y H, Liang C, 2008. Recovery of transgenic plants by pollen-mediated transformation in Brassica juncea. Transgenic Research, 17(3): 417~424
Wantana T. 1994. Genetic transformation in Nicotiana tabacum using pollen as a vector. Kasetsart Journal (Natural Sciences), 28(3): 463~470
Wender P A, Mitchell D J, Pattabiraman K, Pelkey E T, Steinman L, Rothbard J B. 2000. The design, synthesis, and evaluation of molecules that enable or enhance cellular uptake: peptoid molecular transporters. Proceedings of the National Academy of Sciences of the United States of America, 97(24):13003~13008
Widholm J M, Chinnala A R, Ryu J H, Song H S, Egett T, Brotherton J E. 200l. Glyphosate selection of gene amplification in suspension cultures of three plant species. Physiologia Plantarum, l12(4): 540~545
Yang Y S, Watson W J, Tucker P W, Capra J D. 1993. Construction of recombination DNA by exonuclease recession. Nucleic Acids Research, 21(8): 1889~1893
Yoh H, Takeshi Y, Hideo K. 1997. Transformants through pollination of mature maize (zea mays L.) pollen deliverd bar gene by particle gun. Grassland Science, 43(2): 117~123
Zeng J Z, Wang D J, Wu Y Q, Zhang J. 1994. Transgenic wheat plants obtained with pollen-tube pathway method. Science in China (Series B), 37(3): 319~326
Zhou B, Donnelly M E, Scholes D T, George K St, Hatta M, Kawaoka Y, Wentworth D E. 2009. Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and awine origin human InfFluenza a viruses. Journal of Virology, 83(19): 10309~10313
Zhu B G, Cai G F, Hall E O, Freeman G J. 2007. In-FusionTM assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations. BioTechniques, 43(3): 354~359
Zimmermann U, Pilwat G, Pequeux A, Gilles R. 1980. Electro-mechanical properties of human erythrocyte membranes:the pressure-dependence of potassium permeability. Journal of Memberane Biology, 54(2): 103~113
Zorko M, Langel U. 2005. Cell-penetrating peptides: mechanism and kinetics of cargo delivery. Advanced Drug Delivery Reviews, 57(4): 529~545
陈其军,安瑞,周海梦,陈珈,王学臣. 2004.使用Gateway技术兼容的T载体获得入门克隆.生物化学与生物物理进展, 31(10): 951~954
陈茜,刘亚伟,黄邵,杨勤,姜勇. 2009.细胞穿透肽穿膜机制的研究进展.国际病理科学与临床杂志, 29(2): 115~120
邓丽琴,祝朋芳,陈长青. 2004.试论常规育种与分子育种的研究应用.杂粮作物, 24(005): 280~281
丁志山,沃兴德. 1997.超声波的生物学效用及其在转基因中的应用.生命科学, 9(4): 187~189
董云洲,段胜军,赵连元,杨秀海,贾士荣. 1999.用基因枪转化花粉获得转基因谷子和玉米.中国农业科学, 32(2): 9~13
杜春芳,刘惠民,李朋波,孙毅,李润植. 2006.花粉介导法获得油菜转基因植株研究.作物学报, 32(5): 749~754
高东尧,夏兰芹,徐兆师,马有志,李彦舫,原亚萍,李连城,陈明,孙金海. 2009.小麦WVp-1基因表达载体的Gateway技术构建及其遗传转化.麦类作物学报, 29(2): 189~194
赫福霞,郎志宏,陆伟,林敏,张杰,黄大昉. 2008.以耐草甘膦2mG2-epsps基因为选择标记的玉米转化体系的建立.生物技术通报, 5: 92~97.
何迎春,任春梅,高必达. 2001.耐草甘膦基因克隆和作物转化研究新进展.生物技术, 11(4): 38~40
胡金艳,张书祥,王晓武,武剑. 2007.基于Gateway技术的拟南芥雄性不育GHF基因表达载体的构建. 农业生物技术科学, 23(12): 45~48
梁雪莲,郭平毅,孙毅,刘惠民,王景雪,刘少翔,赵志国,唐思静. 2005.玉米3种非组培转基因方法转化外源bar基因研究.作物学报, 31(12): 1648~1653
梁雪莲,郑奕雄,庄东红,黄德强. 2006.花生CpTI基因转化----花粉介导法.花生学报, 35(4): 1~5
林朝贵,范林,陈良龙. 2010.利用In-Fusion技术构建存活素-增强型绿色荧光蛋白融合基因重组慢病毒表达载体.心血管康复医学杂志, 19(1): 10~14
林春晶,韦正乙,蔡勤安,侯敬尧,邢少辰. 2008.几种植物转基因表达载体的构建方法.生物技术, 18 (5): 84~87
刘纪麟. 2001.玉米育种学,北京:中国农业出版社
刘锡娟,刘昱辉,王志兴,王旭静,张永强. 2007.转5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)基因抗草甘膦烟草和棉花的获得.农业生物技术学报, 15(6): 958~963
楼秀芹,周骋. 2008.特洛伊肽运载核酸的研究进展.医学研究杂志, 37(2): 92~95
陆云华,马立新,蒋思婧. 2006.一种通用高效的复杂载体构建的新方法.遗传, 28(2): 212~218
欧阳平,李玉,严红,马立新. 2007.一种载体构建的新方法:一步克隆法.湖北大学学报(自然科学版), 29(2): 178~181
石清琢,姜敏. 2005.玉米转基因育种技术概述.杂粮作物, 25(4): 230~231
苏少泉. 2009.转基因抗草甘膦作物的新进展.现代农药, 8(6): 1~4
孙敬儒,王保义. 2001.强电场脉冲致细胞膜电穿孔的观察.四川大学学报:自然科学版, 38(5):748~751
杨艳芹,谢菊芳,夏利霞,侯义峰,肖磊. 2006.脉冲电场致细胞膜电穿孔的机理分析.湖北大学学报:自然科学版, 28(2): 165~167
万丙良,张献龙. 2000.基因枪介导转化水稻花粉获得转基因植株.农业生物技术学报, 8(4): 332, 381
王宏伟,梁业红,史振声,张世煌. 2007.作物抗草甘膦转基因研究概况术.作物杂志, 4: 9~12
王节之,郝晓芬,郑向阳,王路英,孙毅,王景雪. 2004.谷子花粉介导法转几丁质酶基因的研究.生物技术, 14(5): 5~6
王景雪,孙毅,崔贵梅,胡晶晶. 2001.花粉介导法获得玉米转基因植株.植物学报, 43(3): 275~279
王秀君,郎志宏,陆伟,林敏,单安山,黄大昉. 2008.利用花粉管通道法将耐草甘膦基因mG2-epsps导入玉米自交系的研究初报.中国农业科技导报, 10(4): 56~62
徐化学,熊建华,傅彬英. 2007.应用Gateway技术构建水稻OsDAD1基因的RNA干扰载体.分子植物育种, 5(1): 133~136
张宏,王国英,谢友菊,戴景瑞,许宁,赵南明,李太源,田颖川,乔利亚,莽克强. 1997.超声波介导
转化玉米愈伤组织及可育转基因植株的获得.中国科学(C辑), 27(2): 162~167
张素芝,荣廷昭. 2008.农杆菌介导的玉米遗传转化系统研究进展.遗传, 30(10): 1249~1256
郑秀芳,金治平. 2002.植物遗传转化技术研究概况.河西学院学报, 5: 66~70
周双林,夏苗芬. 2006.肽核酸的研究与应用.中国药学杂志, 41(15): 1124~1128
朱玉. 2002.极端污染环境下草甘膦耐受菌株荧光假单胞菌G2的鉴定及其EPSP合酶基因的克隆. [硕士学位论文].北京:中国农业科学院
朱玉,于中连,林敏. 2003.草甘膦生物抗性和生物降解及其转基因研究.分子植物育种, 1(4): 435~441
Albarran B, To R, Stayton P S. 2005. A TAT-streptavidin fusion protein directs uptake of biotinylated cargo into mammalian cells. Protein Engineering Design and Selection, 18(3): 147~152
Arnizen C J, DeBonte Jr L R, Evans D A, Loh W H, O'dell J T. 1991. 1991-09-17. Pollen-mediated gene transformation in plants. United States Patents, 5049500
Astriab-Fisher A, Sergueev D, Fisher M, Shaw B R, Juliano R L. 2002. Conjugates of antisense oligonucleotides with the Tat and Antennapedia cell-penetrating peptides: effects on cellular uptake, binding to target sequences and biologic actions. Pharmaceutical Research, 19(6): 744~754
Benoit R M, Wilhelm R N, Scherer-Becker D, Ostermeier C. 2006. An improved method for fast, robust, and seamless integration of DNA fragments into multiple plasmids. Protein Expression and Purification, 45(1): 66~71
Berrow N S, Alderton D, Owens R J. 2009. The precise engineering of expression vectors using high-throughput In-Fusion PCR cloning. Methods in Molecular Biology, 498: 75~90.
Berrow N S, Alderton D, Sainsbury S, Nettleship J, Assenberg R, Rahman N, Stuart D I, Owens R J. 2007. A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Research, 35(6): e45
Booy G, Krens F A, Huizing H J. 1989. Attempted pollen-mediated transformation of maize. Journal of Plant Physiology, 135(3): 319~324
Brewbaker J L, Kwack B H. 1963. The essential role of calcium ion in pollen germination and pollen tube growth. American Journal of Botany, 50(9): 859~865
Brooks H, Lebleu B, Vives E. 2005. Tat peptide-mediated cellular delivery: back to basics. Advanced DrugDelivery Reviews, 57(4): 559~577
Chang M, Chou J C, Lee H J. 2005. Cellular internalization of fluorescent proteins via arginine-rich intracellular delivery peptide in plant cells. Plant and Cell Physiology, 46(3): 482~488
Chiu Y L, Ali A, Chu C Y, Cao H, Rana T M. 2004. Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells. Chemistry & Biology, 11: 1165~1175
Chowrira G M, Akella V, Lurquin P F. 1995. Eletroporation-mediated gene transfer into intact nodal meristems in planta: generating transgenic plants without in vitro tissue culture. Molecular Biotechnology, 3(1): 17~23
Chugh A, Amundsen E, Eudes F. 2009. Translocation of cell-penetrating peptides and delivery of their cargoes in triticale microspores. Plant Cell Reports, 28(5): 801~810
Chugh A, Eudes F. 2007. Translocation and nuclear accumulation of monomer and dimer of HIV-1 Tat basic domain in triticale mesophyll protoplasts. Biochimica et Biophysica Acta, 1768(3): 419~426
Chugh A, Eudes F. 2008. Study of uptake of cell penetrating peptides and their cargoes in permeabilized wheat immature embryos. FEBS Journal, 275(10):2403~2414
Curtis M D, Grossniklaus V. 2003. A gateway cloning vector site for high-throughput functional analysis of genes in planta. Plant Physiology, 133(2): 462~469
D’Hallui K, Bonne E, Bossut M, De Beuckeleer M, Leemans J. 1992. Transgenic maize plants by tissue electroporation. Plant Cel1, 4(12): 1495~1505
Dietz G P H, Bahr M. 2007. Synthesis of cell-penetrating peptides and their application in neurobiology. Neuroprotection Methods and Protocols, 399: 181~198
Dill G M. 2005. Glyphosate-resistant crops: history, status and future. Pest Management Science, 61(3): 219~224
Dong J, Kharb P, Teng W, Hall T C. 2001. Characterization of rice transformed via an Agrobacterium-mediated inflorescence approach. Molecular Breeding, 7(3): 187~194
Duke S O. 2005. Taking stock of herbicide-resistant crops ten years after introduction. Pest Management Science, 61(3): 211~218
Fawell S, Seery J, Daikh Y, Moore C, Chen L L, Pepinsky B, Barsoum J. 1994. Tat-mediated delivery of heterologous proteins into cells. Proceedings of the National Academy of Sciences of the United States of America, 91(2): 664~668
Ferrari A, Pellegrini V, Arcangeli C, Fittipaldi A, Giacca M, Beltram F. 2003. Caveolae-mediated internalization of extracellularHIV-1 Tat fusion proteins visualized in real time. Molecular Therapy, 899(2): 284~293
Fittipaldi A, Ferrari A, Zooppe M, Arcangeli C, Pellegrini V, Beltram F, Giacca M. 2003. Cell membrane lipid rafts mediated caveolar endocytosis of HIV-1 Tat fusion proteins. Journal of Biological
Chemistry, 278(36): 34141~34149 Fominaya J, Gasset M, Garcia R, Roncal F, Albar J P, Bernad A. 2000. An optimised amphiphilic cationic peptide as an efficient non-viral gene delivery vector. Journal of Gene Medicine, 2(6): 455~464
Frankel A D, Pabo C O. 1988. Cellular uptake of the tat protein from human immunodeficiency virus. Cell, 55(6): 1189~1193
Fukuoka H, Ogawa T, Mitsuhara I, Iwai T, Isuzigawa K, Nishizawa Y, Gotoh Y, Nishizawa Y, Tagiri A, Ugaki M, Ohshiama M, Yano H, Murai N, Niwa Y, Hibi T, Ohashi Y. 2000. Agrobacterium-mediatedtransformation of monocot and dicot plants using the NCR promoter derived from soybean chlorotic mottle virus. Plant Cell Reports, 19(8): 815~820
Futaki S. 2006. Oligoarginine vectors for intracellular delivery: Design and cellular?\uptake mechanisms. Peptide Science, 84(3): 241~249
Grimsley N, Hohn T, Davies J W, Hohn B. 1987. Agrobacterium-mediated delivery of infectious maize streak virus into maize plants. Nature, 325(8): 177~179
Gordon-Kamm W J, Spencer T M, Mangano M L, Adams T R, Daines R J, Start W G, O’Brien J V, Chambers S A, Adams Jr W R, Willetts N C, Rice T B, Mackey C J, Krueger R W, Kausch A P, Lemaux PG. 1990. Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell, 2(7): 603~618
Gould J, Devey M, Hasegawa O, Ulian E C, Peterson G, Smith R H. 1991. Transformation of Zea mays L. using Agrobacterium tumefaciens and the shoot apex. Plant Physiology, 95(2): 426~434
Gasser C S, Fraley R T. 1989. Genetically Engineering Plants for Crop Improvement. Science, 244(4910): 1293~1299
Green M, Loewenstein P M. 1988. Autonomous functional domains of chemically synthesized human immunodeficiency virus tat trans-activator protein. Cell, 55(6): 1179~1188
Green J M. 2007. Review of glyphosate and ALS-inhibiting herbicide crop resistance and resistant weed management. Weed Technology, 21(2): 547~558
Hallui K, Bonne E, Bossut M, Beuckeleer M D, Leemans J. 1992. Transgenic maize plants by tissue electroporation. Plant Cell, 4(12): 1495~1505
Hamilton D A, Roy M, Rueda J , Sindhu R K, Sanford, Mascarenhas J P. 1992. Dissection of a pollen-specific promoter from maize by transient transformation assays. Plant Molecular Biology, 18(2): 211~218
Hansen G, Wright M S. 1999. Recent advances in the transformation of plants. Trends in Plant Science, 4(6): 226~231
Hartley J L, Temple G F, Brasch M A. 2000. DNA cloning using in vitro site-specific recombination. Genome Research, 10: 1788~1795
Hay L, Lachance D, Vonaderkas P, Charest P J. 1994. Transient chimeric gene expression in pollen of five conifer species following micmprojectile bombardment. Canadian Journal of Forest Research, 24: 2417~2423
Hellgren I, Gorman J, Sylven C. 2004. Factors controlling the efficiency of Tat-mediated plasmid DNA transfer. the Journal of Drug Targeting, 12(1): 39~47
Hess, D., Gresshoff, P.M., Fielitz, U., Gleiss, D. 1974. Uptake of protein and bacteriophage into swelling and germinating pollen of Petunia hybrida. Z. Pflanzenphysiol, 74: 371~376
Hess D, Lorz H. 1974. Uptake of bacterial DNA into swelling and germinating pollen grains of Petunia hybrida and Nicotiana glauca. Z. Pflanzenphysiol, 74: 52~63
Hess D. 1977. Cell modification by DNA uptake. In: Reinert J, Bajaj YPS (eds) Applied and fundamental aspects of plant cell, tissue, and organ culture. Springer-Verlag, Berlin Heidelberg, pp506~577
Hess D. 1980. Investigations on the intra- and interspecific transfer of anthocyanin genes using pollen as vectors. Z Pflanzenphysiol, 98: 321~337
Hess D. 1987. Pollen-based techniques in genetic manipulation. International Reciew of Cytology, 107:367~395
Ishida Y, Saito H, Ohta S, Hiei Y, Komari T, Kumashiro T. l996. High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens. Nature Biotechnology, l4(6): 745~750
Jacob G S, Garbow O R, Hallas L E, Kimack N M, Kishore G M Schaefer J.1988. Metabolism of Glyphosate in Pseudomonas sp. Strain LBr. Applied and Environmental Microbio1ogy, 54(12):
2953~2958 Jeang K T, Xiao H, Rich E A. 1999. Multifaceted activities of the HIV-1 transactivator of transcription, Tat. the Journal of Biological Chemistry, 274(41): 28837~28840
Joersbo M, Brunstedt J. 1990. Direct gene transfer to plant protoplasts by mild sonication. Plant Cell Reports, 9(4): 207~210
Karimi M, Depicker A and Hilson P. 2007. Recombinational cloning with plant gateway vectors. Plant Physiology, 145(4): 1144~1154
Klein T M, Fromm M, Weissinger A, Tomes D, Schaaf S, Sletten M, Sanford J C. 1988. Transfer of foreign genes into intact maize cells with high-velocity microprojectiles. Proceedings of the National Academy of Sciences of the United States of America, 85(12): 4305~4309
Koppelhus U, Awasthi S K, ZacharV, Holst H U, Ebbesen P, Nielsen P E. 2002. Cell-dependent differential cellularuptake of PNA, peptides and PNA-peptide conjugates. Antisense Nucleic Acid Drug Development, 12(2): 51~63
Landy A. 1989. Dynamic, structural and regulatory aspects ofλsite-specific recombination. Annual Reviews of Biochemistry, 58: 913~941
Lansac A R, Sullivan C Y, Johnson B E, Lee K W. 1994. Viability and germination of the pollen of sorghum [Sorghum bicolor (L.) Moench]. Annals of Botany, 74(1): 27~33
Li Y H, Tremblay F M, Séguin A. 1994. Transient transformation of pollen and embryogenic tissues of white spruce (Picea glauca(Moench.) Voss) resulting from microprojectile bombardment. Plant Cell Reports, 13(12): 66l~665
Lindsay M A. 2002. Peptide-mediated cell delivery: application in protein target validation. Current Opinion in Pharmacology, 2(5): 587~594
Loret E P, Vives E, Ho P S, Rochat H, Van Rietschoten J, Johnson Jr W C. 1991. Activating region of HIV-1 Tat protein: vacuum UV circular dichroism and energy minimization. Biochemistry, 30(24): 6013~6023
Marsischky G, LaBaer J. 2004. Many paths to many clones: a comparative look at high-throughput cloning methods. Genome Research, 14: 2020~2028.
Martinussen I, Junttila O, Twell D. 1994. Optimization of transient gene expression in pollen of Norway spruce (Picea abies) by particle acceleration. Physiologia Plantarum, 92(3): 412~416
Matthews B F, Abdul-Baki A A, Saunders J A. 1990. Expression of a foreign gene in electroporated pollen grains of tobacco. Sexual Plant Reproduction, 3(3): 147~151
McCormick S, Yamaguchi J, Twell D. 1991. Deletion analysis of pollen-expressed promoters. In Vitro Cellular & Developmental Biology-Plant, 27(1): 15~20
McCabe D, Christou P. 1993. Direct DNA transfer using electric discharge particle acceleration (ACCELLTM technology). Plant Cell, Tissue and Organ Culture, 33(3): 227~236
Nafziger E D, Widholm J M, Steinrucken H C, Killmer J L. 1984. Selection and characterization of a carrotcell line tolerant to glyphosate. Plant Physiology, 76(3): 571~574
Nishihara M, Ito M, Tanaka l, Kyo M, Ono K, Irifune K, Morikawa H. 1993. Expression of theβ-glucuronidase gene in pollen of lily(Lilium longiflorum),tobacco (Nicotiana tabacum),Nicotiana rustica,and peony (Paeonia lactiflora)by paricle Bombardment. Plant Physiology, 102: 357~361
Ohta Y. 1986. High-efficiency genetic transformation of maize by a mixture of pollen and exogenons DNA. Proceedings of the National Academy of Sciences of the United States of America, 83: 715~719
Penaloza A, Mena G L, Herrera L, Bailey A M. 1995. Cloning and sequencing of the genes involved in glyphosate utilization by Pseudomonas pseudomallei. Applied Environmental Microbiology, 61(2): 538~543
Potrykus I. 1990. Gene transfer to plants: assessment and perspectives. Physiologia Plantarum, 79(1): 125~134
Rhodes C A, Pierce D A, Mettler I J, Mascarenhas D, Detmer J J. 1988. Genetically transformed maize plants form protoplasts. Science, 240(4849): 204~207
Ruben S, Perkins A, Purcell R, Joung K, Sia R, Burghoff R, Haseltine W, Rosen C. 1989. Structural and functional characterization of human immunodeficiency virus tat protein. Journal of Virology, 63(1): 1~8
Rudolph C, Planks C, Lausier J, Schillinger U, Muller R H, Rosenecker J. 2003. Oligomers of the arginine-rich motif of the HIV-1TAT protein are capable of transferring plasmid DNA into cells. the Journal of Biological Chemistry. 278(13): 11411~11418
Saalik P, Elmquist A, Hansen M, Padari K, Saar K, Viht K, Langel U, Pooga M. 2004. Protein cargo delivery properties of cell-penetrating peptides. A comparative study. Bioconjugate Chemistry, 15(6): 1246~53
Sanford J C, Skubik K. 1986. Attempted pollen-mediated transformation using Ti-plasmids. Springer-Verlag, New York (USA), US8719725
Sanford J C, Klein T M, Wolf E D, Allen N. 1987. Delivery of substances into cells and tissues using a particle bombardment process. Particulate Science and technology, 5(1): 27~37
Sanford J C , Skubik K A , Reishch B I. 1985. A ttempted pollen-mediated plant transformation employing genomic donor DNA. Theoretical and Applied Genetics, 69(5-6): 571~574
Sasaki Y, Sane T, Yoshida S, Yahata K, Hotta T, Chesnut J D, Honda T, Imamoto F. 2004. Evidence for high specificity and efficiency of multiple recombination signals in mixed DNA cloning by the Multisite Gateway system. Journal of Biotechnology, 107(3): 233~243
Schwarze S R, Dowdy S F. 2000. In vivo protein transduction: intracellular delivery of biologically active proteins, compounds and DNA. Trends in Pharmacological Sciences, 21(2): 45~48
Sidorov V, Gilbertson L, Addae P, Duncan D. 2006. Agrobacterium-mediated transformation of seedling-derived maize callus. Plant Cell Reports, 25(4): 320~332
Sleight S C, Bartley B A, Lieviant J A, Sauro H M. 2010. In-Fusion BioBrick assembly and re-engineering. Nucleic Acids Research, 38 (8): 2624~2636
Smith C R, Saunders J A, Wer S V, Cheng J P, Matthews B F. 1994. Expression of GUS and CAT activities using electrotransformed pollen. Plant Science, 104(1): 49~58
Stoger E, Moreno R M B, Ylstra B, Vicente O, Heberle-Bors E. 1992. Comparison of different techniques for gene transfer into mature and immature tobacco pollen. Transgenic Research, 1(2): 71~78
Tan S, Evans R, Singh B. 2006. Herbicidal inhibitors of amino acid biosynthesis and herbicide-tolerant crops. Amino Acids, 30(2): 195~204
Tenno T, Goda N, Tateishi Y, Tochio1 H, Mishima M, Hayashi H, Shirakawa M, Hiroaki H. 2004. High-throughput construction method for expression vector of peptides for NMR study suited for isotopic labeling. Protein Engineering, Design & Selection, 17(4): 305~314
Torchilin V P, Levchenko T S, Rammohan R, Volodina N, Papahadjopoulos-Sternberg B, D’Souza G G M. 2003. Cell transfection in vitro and in vivo with non-toxic TAT peptide-liposome-DNA complexes. Proceedings of the National Academay of Sciences of the United States of the America, 100(4): 1972~1977
Torchilin V P, Rammohan R, Weissig V, Levchenko T S. 2001. Tat peptide on the surface of liposomes sffords their efficient intracellular delivery even at low temperature and in the presence of metabolic inhibitors. Proceedings of the National Academy of Sciences of the United States of America, 98(15): 8786~8791
Tung C H, Mueller S, Weissleder R. 2002. Novel branching membrane translocational peptide as gene delivery vector. Bioorganic & Medicinal Chemistry, 10(11): 3609~3614
Turner R , Bate N, Twell D, Foster G D. 1994. In vivo characterisation of a translational enhancer upstream from the coat protein open reading frame of potato virus S. Archives of Virology, 137: 123~132
Twell D, Klein T M., Fromm M E., McCormick S. 1989. Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiology, 91(4): 1270~1274
Twell D, Patel S, Sorensen A, Roberts M, Scott R, Draper J, Foster G. 1993. Activation and developmental regulation of an Arabidopsis anther-specific promoter in micmpores and pollen of Nicotiana tabacum. Sexual Plant Reproduction, 6: 217~224
Unnamalai N, Kang B G, Lee W S. 2004. Cationic oligopeptide-mediated delivery of dsRNA for post-transcriptional gene silencing in plant cells. FEBS Letters, 566(1-3): 307~319
van der Leede-Plegt L M, van de Ven B C E, Bino R J, van der Salm T P M, van Tunen A. 1992. Introduction and differential use of various promoters in pollen grains of Nicotiana glutinosa and Lilium longiflorum. Plant Cell Reports, 11(1): 20~24
van der Leede-Plegt L M , van de Ven B C E , Schilder M, Franken J, van Tunen A J. 1995. Development of a pollen-mediated transformation method for Nicotiana glutinosa. Transgenic Research, 4(2): 77~86
Vivès E, Brodin P and Lebleu B. 1997. A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. Journal of Biological Chemistry, 272(25): 16010~16017
Wadia J S, Dowdy S F. 2002. Protein transduction technology. Current opinion in biotechnology, 13(1): 52~56
Wang J X, Li Y H, Liang C, 2008. Recovery of transgenic plants by pollen-mediated transformation in Brassica juncea. Transgenic Research, 17(3): 417~424
Wantana T. 1994. Genetic transformation in Nicotiana tabacum using pollen as a vector. Kasetsart Journal (Natural Sciences), 28(3): 463~470
Wender P A, Mitchell D J, Pattabiraman K, Pelkey E T, Steinman L, Rothbard J B. 2000. The design, synthesis, and evaluation of molecules that enable or enhance cellular uptake: peptoid molecular transporters. Proceedings of the National Academy of Sciences of the United States of America, 97(24):13003~13008
Widholm J M, Chinnala A R, Ryu J H, Song H S, Egett T, Brotherton J E. 200l. Glyphosate selection of gene amplification in suspension cultures of three plant species. Physiologia Plantarum, l12(4): 540~545
Yang Y S, Watson W J, Tucker P W, Capra J D. 1993. Construction of recombination DNA by exonuclease recession. Nucleic Acids Research, 21(8): 1889~1893
Yoh H, Takeshi Y, Hideo K. 1997. Transformants through pollination of mature maize (zea mays L.) pollen deliverd bar gene by particle gun. Grassland Science, 43(2): 117~123
Zeng J Z, Wang D J, Wu Y Q, Zhang J. 1994. Transgenic wheat plants obtained with pollen-tube pathway method. Science in China (Series B), 37(3): 319~326
Zhou B, Donnelly M E, Scholes D T, George K St, Hatta M, Kawaoka Y, Wentworth D E. 2009. Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and awine origin human InfFluenza a viruses. Journal of Virology, 83(19): 10309~10313
Zhu B G, Cai G F, Hall E O, Freeman G J. 2007. In-FusionTM assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations. BioTechniques, 43(3): 354~359
Zimmermann U, Pilwat G, Pequeux A, Gilles R. 1980. Electro-mechanical properties of human erythrocyte membranes:the pressure-dependence of potassium permeability. Journal of Memberane Biology, 54(2): 103~113
Zorko M, Langel U. 2005. Cell-penetrating peptides: mechanism and kinetics of cargo delivery. Advanced Drug Delivery Reviews, 57(4): 529~545