“睡美人”转座子系统对于外源基因整合和表达促进作用
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摘要
“睡美人”(Sleeping Beauty , SB)转座系统属于Tc1/mariner转座子家族,已失活了一千多万年。1997年Ivics等根据积累的系统发生数据,利用生物信息学的方法进行分子重建,终于使其恢复转座活性。近年来对“睡美人”转座系统转座效率和转座机理的研究表明SB转座子系统在转基因,基因筛选,促进外源基因表达及基因治疗等领域具有广阔的应用前景。
本研究利用“睡美人”转座子系统,比较睾丸直接注射法和SB系统介导下睾丸注射法转基因小鼠的制备效率,并分析了SB系统中的转座酶基因对基因免疫的增强作用。首先从本实验室已构建的精囊腺组织特异性表达载体(PC-EGFP)切出8.4kb含有EGFP的cDNA及其5’和3’调控序列表达片段,插入到“睡美人”转座子载体PT2-HB中获得PT2-EGFP。随后对睾丸直接注射制备转基因小鼠方法进行初步优化,利用已构建的SB系统和经过双酶切PC-EGFP获得的8.4kbDNA片段分别进行睾丸注射制备转基因动物,利用PCR方法比较转基因动物的制备效率,并对“睡美人”转座子与转座酶的比例进行了初步的摸索。研究结果表明,用4号针头,进行3次注射的公鼠与野生型ICR母鼠合笼后代的产子数及转基因阳性率较理想;经PCR并结合精囊腺和阴道栓的表达检测,SB系统介导的转基因阳性率(43%左右)显著高于普通8.4kb DNA片段的注射组(20%左右),“睡美人”转座子与转座酶的比例在5:1-20:1范围,后代的阳性率差异不显著。
本试验同时构建了表达人乳铁蛋白(human lactoferrin,hLF)的真核表达载体pA-hLF,在进行家兔人乳铁蛋白多抗的制备过程中,按照“睡美人”转座酶:PA-LF=4:1的比例用PEI包裹后注射新西兰家兔,并设置PA-LF与PEI包裹,裸露的PA-LF两个注射组与对照组,以评价“睡美人”转座酶对于外源基因表达的促进作用。结果表明:“睡美人”转座酶与PA-LF注射组其抗体效价可达1:6400,高于PEI包裹PA-LF组(1:1600)和裸露质粒注射组(1:2000)。研究结果表明“睡美人”转座酶对基因免疫具有促进作用,可作为一种新型基因佐剂在抗体制备过程中发挥一定的作用。
The Sleeping Beauty (SB) transposon is a member of Tc1/mariner family transposons that has been transpositionally inactive for over 10 million years. The SB transposon system was awakened from inactive Tc1-like transposable elements by using molecular phylogenetic data in 1997. Recent studies on its transposition efficiency and mechanism have shown its broad applications in vertebrate animals for gene-screening, gene transfer, and human gene therapy.
In this study, the trangenic efficiency introduced by SB system in mouse and effect on gene immunization of Sleeping Transposase was analysed. Firstly, method of injection was optimized by exlporing the size of injector pinhead, injecting times and hormone in this research. The vector PT2-EGFP was constructed by inserting 8.4kb fragment including EGFP and its 5’and 3’regulated sequence from constructed seminal vesicle vector(PC-EGFP) into PT2-HB. Transgenic animals were obtained by direct testis injection and injection mediated by SB transposon system respectively.The transgenic rate were evaluated and compared between these two methods by detection of the fluorescence of vaginal plug and section of seminal vesicle under fluorescence microscope and PCR. In addtion, the different ratio of tansposon and tansposase in SB transposon system was assessed for high transgenic rate. The results demonstrated that the transgenic rate and litter size of offspring in the groups injected by NO.4 pinhead and injected three times were higher. The transgenic rate mediated by SB(43%) was higher than the transgenic rate by direct testisinjection(20%). However, the transgenic rate was not significant different among the groups with different ratio of transposon and transposase from 5:1-20:1. The results demontrated that the Sleeping Beauty system can enhance the transgenic rate.
To investigate the effect of Sleeping Beauty transposase on gene immunazition, the vector pA-hLF was constructed and expressed lactoferritin for gene immunization in Newsland rabbits. 8 weeks old rabbits were divided into four groups: group 1, nake vector-pA-hLF;group 2, PEI-coated pA-hLF;group 3, PEI coated complex with Sleeping Transposase and pA-hLF at a molar ratio of 4:1,;group 4, the control group withough injection.Then intrmuscula immunizations were conducted every two weeks in first 6 week and the final injection was given in 18th week. the blood was sampled and evaluated the titer. After the fourth immunizations the third group produce the antibody against hLF by 1:6400 higher than the first group (1:1600) and the second group (1:2000), and the control group showed no antibody. The result indicate that SB tansposase can promote the expression of exogenous gene, which could be used as a new adjuvant in gene immunization.
本研究利用“睡美人”转座子系统,比较睾丸直接注射法和SB系统介导下睾丸注射法转基因小鼠的制备效率,并分析了SB系统中的转座酶基因对基因免疫的增强作用。首先从本实验室已构建的精囊腺组织特异性表达载体(PC-EGFP)切出8.4kb含有EGFP的cDNA及其5’和3’调控序列表达片段,插入到“睡美人”转座子载体PT2-HB中获得PT2-EGFP。随后对睾丸直接注射制备转基因小鼠方法进行初步优化,利用已构建的SB系统和经过双酶切PC-EGFP获得的8.4kbDNA片段分别进行睾丸注射制备转基因动物,利用PCR方法比较转基因动物的制备效率,并对“睡美人”转座子与转座酶的比例进行了初步的摸索。研究结果表明,用4号针头,进行3次注射的公鼠与野生型ICR母鼠合笼后代的产子数及转基因阳性率较理想;经PCR并结合精囊腺和阴道栓的表达检测,SB系统介导的转基因阳性率(43%左右)显著高于普通8.4kb DNA片段的注射组(20%左右),“睡美人”转座子与转座酶的比例在5:1-20:1范围,后代的阳性率差异不显著。
本试验同时构建了表达人乳铁蛋白(human lactoferrin,hLF)的真核表达载体pA-hLF,在进行家兔人乳铁蛋白多抗的制备过程中,按照“睡美人”转座酶:PA-LF=4:1的比例用PEI包裹后注射新西兰家兔,并设置PA-LF与PEI包裹,裸露的PA-LF两个注射组与对照组,以评价“睡美人”转座酶对于外源基因表达的促进作用。结果表明:“睡美人”转座酶与PA-LF注射组其抗体效价可达1:6400,高于PEI包裹PA-LF组(1:1600)和裸露质粒注射组(1:2000)。研究结果表明“睡美人”转座酶对基因免疫具有促进作用,可作为一种新型基因佐剂在抗体制备过程中发挥一定的作用。
The Sleeping Beauty (SB) transposon is a member of Tc1/mariner family transposons that has been transpositionally inactive for over 10 million years. The SB transposon system was awakened from inactive Tc1-like transposable elements by using molecular phylogenetic data in 1997. Recent studies on its transposition efficiency and mechanism have shown its broad applications in vertebrate animals for gene-screening, gene transfer, and human gene therapy.
In this study, the trangenic efficiency introduced by SB system in mouse and effect on gene immunization of Sleeping Transposase was analysed. Firstly, method of injection was optimized by exlporing the size of injector pinhead, injecting times and hormone in this research. The vector PT2-EGFP was constructed by inserting 8.4kb fragment including EGFP and its 5’and 3’regulated sequence from constructed seminal vesicle vector(PC-EGFP) into PT2-HB. Transgenic animals were obtained by direct testis injection and injection mediated by SB transposon system respectively.The transgenic rate were evaluated and compared between these two methods by detection of the fluorescence of vaginal plug and section of seminal vesicle under fluorescence microscope and PCR. In addtion, the different ratio of tansposon and tansposase in SB transposon system was assessed for high transgenic rate. The results demonstrated that the transgenic rate and litter size of offspring in the groups injected by NO.4 pinhead and injected three times were higher. The transgenic rate mediated by SB(43%) was higher than the transgenic rate by direct testisinjection(20%). However, the transgenic rate was not significant different among the groups with different ratio of transposon and transposase from 5:1-20:1. The results demontrated that the Sleeping Beauty system can enhance the transgenic rate.
To investigate the effect of Sleeping Beauty transposase on gene immunazition, the vector pA-hLF was constructed and expressed lactoferritin for gene immunization in Newsland rabbits. 8 weeks old rabbits were divided into four groups: group 1, nake vector-pA-hLF;group 2, PEI-coated pA-hLF;group 3, PEI coated complex with Sleeping Transposase and pA-hLF at a molar ratio of 4:1,;group 4, the control group withough injection.Then intrmuscula immunizations were conducted every two weeks in first 6 week and the final injection was given in 18th week. the blood was sampled and evaluated the titer. After the fourth immunizations the third group produce the antibody against hLF by 1:6400 higher than the first group (1:1600) and the second group (1:2000), and the control group showed no antibody. The result indicate that SB tansposase can promote the expression of exogenous gene, which could be used as a new adjuvant in gene immunization.
引文
[1] Mc.Clintock B. Chromosome organization and gene expression .Cold Spring Harbor Symp Quant Biol 1951, 16(3): 13-17.
[2] Yant S. R., Meuse L., Chiu W., et al. Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system. Nat Genet, 2000, 25(1): 35-41.
[3] Fischer S. E., Wienholds E., Plasterk R. H. Regulated transposition of a fish transposon in the mouse germ line. Proc Natl Acad Sci USA, 2001,98(12): 6759-6764.
[4] Sheng D., Hui W. X., Li G., et al. Efficient transposition of the piggyBac resource (PB) transposon in mammalian cells and mice. Cell, 2005, 122(3): 473-483.
[5] Ivics Z., Hackett P. B., Plasterk R. H.,et al. Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Cell, 1997, 91(4): 501-510.
[6] Horie K., Yusa K., Yae K., et al. Characterization of Sleeping Beauty transposition and its application to genetic screening in mice. Mol Cell Biol, 2003, 23(24): 9189-9207.
[7] Izsvak Z., Ivics Z,, Plasterk R. H. Sleeping Beauty, a wide host-range transposon vector for genetic transformation in vertebrates. Mol Biol, 2000, 302: 93-102.
[8] Liu L., Mah C., Fletcher B. S. Sustained FVIII expression and phenotypic correction of Hemophilia A in neonatal mice using an endothelial-targeted Sleeping Beauty transposon. Mol Ther, 2006, 13(5): 1006-1015.
[9]丁昇,吴晓晖.“睡美人”转座系统研究进展.生物化学与生物物理进展,2003,30(1):43-48.
[10] Baus J., Liu L., Arnold D., et al.Hyperactive transposase mutants of the Sleeping Beauty Transposon. Mol Ther, 2005, 12(6): 1148-1156.
[11] Cui Z., Geurts A. M., Liu G., et al.Structure-function analysis of the inverted terminal repeats of the sleeping beauty transposon. Mol Biol, 2002, 318(5): 1221-1235.
[12] Geurts A. M., Yang Y., Clark K. J., et al. Gene transfer into genomes of human cells by the Sleeping Beauty transposon system. Mol Ther, 2003, 8(1): 108-117.
[13] Zayed H., Izsvak Z., Khare D., et al.The DNA bending protein HMGB1 is a cellular cofactor of Sleeping Beauty transposition. Nucleic Acids Res, 2003, 31(9): 2313-2322.
[14] Dupuy A. J., Fritz S., Largaespada D. A. Transposition and gene disruption in the male germline of the mouse. Genesis, 2001, 30(2): 82-88.
[15] Horie K., Kuroiwa A., Ikawa M., et al. Efficient chromosomal transposition of a Tc1/mariner- like transposon Sleeping Beauty in mice. Proc Natl Acad Sci USA, 2001, 98(16): 9191-9196.
[16] Doak T. G., Doerder F. P., Jahn C. L., et al. A proposed superfamily of transposase genes:transposon-like element in ciliated protozoa and a common“D35E”motif. Proc Natl Acad Sci USA, 1994, 91(3): 942-946.
[17] Masuda K., Yamamoto S., Endoh M., et al. Transposon independent increase of transcription by the Sleeping Beauty transposase.Biochem. Biophys Res Commun, 2004, 317(3): 796-800.
[18] Vigdal T. J., Kaufman C. D., Izsvak Z., et al.Common physical properties of DNA affecting target site selection of Sleeping Beauty and other Tc1/mariner transposable elements. Mol Biol, 2002, 323(3): 441-452.
[19] Izsvak Z., Khare D., Behlke J., et al. Involvement of a bifunctional, paired-like DNA-binding domain and a transpositional enhancer in Sleeping Beauty transposition. Biol Chem, 2002, 277(37): 34581-34588.
[20] Liu L., Sanz S., Heggestad A. D., et al.Endothelial targeting of the Sleeping Beauty transposon within lung. Mol Ther, 2004, 10(1): 97-105.
[21] Izsvak Z., Stuwe E. E., Fiedler D., et al.Healing the wounds inflicted by Sleeping Beauty transposition by double-strandbreak repair in mammalian somatic cells. Mol Cell, 2004, 13(2): 279-290.
[22] Luo G., Ivics Z., Izsvak Z., et al. Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. Proc Natl Acad Sci USA, 1998, 95(18): 10769-10773.
[23] Liu G., Cui Z., Aronovich E. L., et al.Excision of Sleeping Beauty transposons: parameters and applications to gene therapy. Gene Med, 2004, 6(5): 574-583.
[24] Yant S. R., Wu X., Huang Y., et al. High-resolution genome-wide mapping of transposon integration in mammals. Mol Cell Biol, 2005, 25(6): 2085-2094.
[25] Geurts A. M., Hackett C. S., Bell J. B., et al. Structure-based prediction of integration-site preferences of transposons into chromosomes.Nucleic Acids Res, 2006, 34(9): 2803-2811.
[26] Miller D. G., Rutledge E. A., Russell D. W. Chromosomal effects of adeno-associated virus vector integration. Nat Genet, 2002, 30(22): 147-148.
[27]肖巧学,曹阳,黄自然,刘良式.昆虫中的mariner类转座子.遗传, 2000, 22(2): 114-118.
[28] Wilson M. H., Kaminski J. M., George A. L. "Functional zinc finger/sleeping beauty transposase chimeras exhibit attenuated overproduction inhibition." FEBS Lett, 2005, 579(27): 6205-6209.
[29] Yusa K Takeda J, Horie K. Enhancement of Sleeping Beauty transposition by CpG methylation: possible role of heterochromatin formation. Mol Cell Biol, 2004, 24(9): 4004-4018.
[30] Park C. W., Park J., Kren B. T., et al. Sleeping Beauty transposition in the mouse genome is associated with changes in DNA methylation at the site of insertion. Genomics, 2006, 88(2): 204-213.
[31] Hacein B. A., Svon K. C., Schmidt M., et al. LMO2-associated clonal T cellproliferation in two patients after gene therapy for SCID-X1. Science, 2003, 302(5644): 415-419.
[32] Dupuy A .J., Clark K., Carlson C. M., et al.Mammalian germ-line transgenesis by transposition. Proc Natl Acad Sci USA, 2002, 99(7): 4495-4499.
[33] Wilber A. C., Frandsen J. L., Geurts J. L, et al. RNA as a source of transposase for Sleeping Beauty mediated gene insertion and expression in somatic cells and tissues. Mol Ther, 2006, 13(3): 625-630.
[34] Score P. R., Belur L. R., Frandsen J. L., et al. Sleeping Beautymediated transposition and long-term expression in vivo: use of the Cre-LoxP recombinase system to distinguish transposition-specific expression. Mol Ther, 2006, 13(3): 617-624.
[35] Chen Z. J., Kren B. T., Wong P. P., et al. Sleeping Beauty-mediated down-regulation of huntingtin expression by RNA interference. Biochem Biophys Res Commun, 2005, 329(2): 646-652.
[36] Yant S. R., Ehrhardt A., Mikkelsen J. G., et al. Transposition from a gutless adeno-transposon vector stabilizes transgene expression in vivo. Nat Biotech, 2002, 20(10): 999-1005.
[37] Yae K., Keng V. W., Koike M., et al. Sleeping Beauty transposon-based phenotypic analysis of mice: lack of arpc3 results indefective trophoblast outgrowth. Mol Cell Biol, 2006, 26(16): 6185-6196.
[38] Dupuy A . J., Jenkins N. A., Copeland N. G. Sleeping Beauty: a novel cancer gene discovery tool. Hum Mol Genet, 2006, 15(Suppl. 1): R75-R79.
[39] Gordon J. W., Scangos G. A ., P lotkin D. J., et al. Genetic transformation ofmouse embryo by m icro injection of purified DNA. P roc Natl Acad Sci U SA , 1980, 77: 7280- 7384.
[40] Palmiter R. D., Brinster R. E., HammerM. E., et al. D ramatic growth of m ice that developed from eggs micronjected with metallothionein-grow th hormone fusiongenes. Nature, 1982, 300: 611- 615.
[41]庞有志,赵淑娟.转基因技术与鸡的抗病育种.中国兽医科技, 1999, 3: 43- 44.
[42]桑润滋主编.动物繁殖生物技术.北京:中国农业出版社2002.
[43] Chan A. W. S., Homan E. J., Ballou L. U., et al. Transgenic cattle p roduced by reverse-transcribed gene transfer in oocytes. Proc Natl Acad Sci U SA , 1998, 95: 14082 -14033.
[44] Chan A. W. S., Chong K. Y., M artinovich C., et al. T ransgenic monkey p roduced by retroviral gene transfer into mature oocytes. Science, 2001, 291: 309- 312.
[45]成勇,成国祥,张靖溥等.牛As1-酪蛋白-乙肝病毒表面抗原融和基因在转基因羊中的表达.生物工程学报, 1997,13 (2) : 154-159.
[46]崔保安,王莉娟,赵楠等.转基因动物关键技术及其进展.河南畜牧兽医, 2003, (4) : 10-11.
[47]严泉剑.动物转基因与核移值技术的新进展.动物医学进展2002, 23 (3) : 24-29.
[48]杨章平,常洪.转基因动物的研究进展.世界农业, 2001, (3) :39-42.
[49] Janeisch R., Mintz B. Simian virus 40 DNA sequences in DNA healthy adultm ice derived from preimplantation blastocysts injected with viral DNA. P roc N atl A cad Sci U SA , 1974, 71 (4) :1250.
[50]刘谦,朱鑫泉.生物安全.北京:科学出版社, 2001.
[51] Schnieke A. E., Kind A. J., R itchie W. A ., et al. Human facto r IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblaatsp. Science, 1997, 278: 2130-2133.
[52]沈子龙,廖建民,徐寒梅.转基因动物技术与转基因动物制药.中国药科大学学报, 2002, 33 (2) : 81-86.
[53] Brakette B. G., Braransca W., Sawicki W. Uptake of heterologous genome by mammalian spermatozoa and its transfer to ova through fertilization. Proc. Natl. Acad. Sci. USA. ,1971 ,68 (2) :353-357.
[54] Arezzo F. Sea urchin sperm as a vector of foreign genetic information. Cell.Biol. Int . Rep. 1989 , 13 (4) :391 - 404.
[55] Lavitrano M., Camaioni A., Fazio V. M., et al . Sperm cells as vectors for introducing foreign DNA into eggs. Genetic transformation of mice. Cell , 1989 ,57 :717-723.
[56] Brackett B. G., Baranska W., Sawicki W., et al. Up take of hetero logous genome by mammallan spermatozoa and its transfer to ova through fertlization. P roc N atl A cad SciU SA , 1971, 68: 353-357.
[57] Anthony C. F., Teruhiko W. Mammali transgenesis by intracyto plasmic sperm injection. Science, 1999, 284: 1180-1183.
[58] Kim J. H., Jung H. H., Lee H. T. Development of a posi-mousetive method for male stem cell-mediated gene transfer in pig.Mol Reprod Dev.1997 , 46 (4) :515-526.
[59] Yamaza K.Y., Fujimoto H., Ando H. In vivo gene, acid transfer to mouse spermatogenic cells by deoxyribonucleic, acid injection into seminiferous tubules and subsequent elect roporation . Biol Reprod ,1998 ,59 (6) :1439-21444.
[60]沈新民,乔贵林,张玲.用曲细精管微注射法建立绿色荧光蛋白,转基因小鼠.第一军医大学学报.2002 , 22 (3) :250-2253.
[61] Garrett F. E., Goel S., Yasu l. J., et al . Liposomes fuse, impermeant with sperm cells s and induce activation by delivery of agents. Biochim Biophys Acta 1999 , 72 (3) :435-444.
[62]肖红卫,郑新民,陈思怀等.精子介导生产转基因猪.华中农业大学学报.2006,25(2):170-173.
[63] HE X., QI B., LIU G. S., YU W. D., et al. A Novel Method to , Transfer Gene In vivo System. Progress in Biochemistry and Biophysics 2006, 33(7): 685-690.
[64] Atkinson P. W., Hines E. R., Beaton S., et al. Association of exoigenous DNA with cattle and insect spermatozoa in vitro. Mo l Rep rod , 1991, 29 (1) : 1-5.
[65] Lavitrano M.,French D., Zani M., et al. The interaction between exogenousDNAand sperm cells. Mo l Reprod, 1992, 31: 161-169.
[66]翟少伟,谷艳钗.转基因动物的研究进展和前景展望.当代畜牧, 2000, (6) : 123.
[67] Arezzo F. Sea urchin sperm as a vector of foreign genetic informatiom . Cell Biol Int Reprod 1989 , 13 :391 - 404.
[68] Nakanishi A., IritaniA. Gene reansfer in the ch icken by sperm2mediated methods. Mo l Repo rd, 1993,36 (2) : 258-261.
[69] Ebert K. M., Selgrath J. P. Transgenic production of a variant of human tissue-type plasm inogen activator in goat milk: generation of transgenic goats and analysis of expression. Bio techno logy,1991, (9) : 835-838.
[70] Bach iller D., Schellander K., Peli J., et al. L iposome Mediated DNA up take by sperm. Mo l Rep rod,1991, 30 (3) : 194-200.
[71] Wolff J. A., Malone R. W., Williams P., et al. Direct gene transfer into mouse muscle in vivo. Science ,1990 ,247∶1465 -1468.
[72] Hartikka J., Sawdey M., Jensen F. C.,et al. An improved plasmid DNA expression vector for direct inection into skeletal muscle. HumanGene Therapy ,1996 ,7∶1205 -1209.
[73] Benvenisty B., Reshef. Direct introduction of genes into rats and expression of the genes. P roc Natl Sci USA. 1986 12, 83 (24) : 9551-9555.
[74] Guyla A., George D., Donald R. Human dystroph in expression in mdx mice after intramuscular injection of DNA constructs. Natrue, 1991, 352: 815-818.
[75] Tang C., Devit M., Johnston S. A. Genetic immunization is a simple method for eliciting an immune response. Nature,1992, 356 (6365) : 152-154.
[76] Ulmer J. B., Donnelly J. J., Parker S. E., et al. Hetero logous protection against influenza by injection of DNA encoding a viral protein. Science, 1993, 259 (5102) : 1745-1749.
[77]王世若.当前研制预防用疫苗的发展趋势.预防兽医学进展2000 , 30(3)∶15.
[78]解洪业.新型疫苗的研究与发展.青海畜牧兽医杂志1997 , (3)∶24—26.
[79] Schrijver R., Langendijk J. P., Keil G.M, et al. Immunization ofcattle with a BHVi vector vaccine or a DNA vaccine both coding for the G-protein of BR.Vaccine ,1997 ,15∶1908-1916.
[80] Krieg A.M., YiA K., Matson S., et al. CpG motifs in bacterial DNA trigger direct B cell activation. Nature, 1995, 374(6522) : 546-569.
[81] Temperton N. J., Quenelle D.C., Lawson K.M., et al. Enhancement of humoral immune responses to a human cytome galovirus DNA vaccine: Adjuvant effects of aluminum phosphate and CpG oligodeoxynucleo tides. J Med V iro l,2003, 70 (1) : 86-90.
[82] Youssef S., Wildbaum G., Maor G., et al. Long lasting protective immunity to experimental auto immune encephalo myelitis following vaccination with naked DNA encoding C-C chemokines. J Immunol, 1998, 161 (8) : 3870-3879. .
[83] Okada E. SasakiS, Ish ii N., et al. Intranasal immunization of a DNA vaccine with IL -12 and granulocyte macrophage colony stimulating factor (GM-CSF) expressing plasmids in liposomes against HIV-1 antigens. J. Immunol, 1997, 159 (70) : 3638-3647.
[84] Kusakabe K., Xin K. Q., Katoh H., et al. The timing of GM-CSF expression plasmid administration influences the Th1?T h rsponse induced by an H IV -1 specific DNA vaccine. J.Immuno l, 2000, 164 (6) : 3102-3111.
[85] Iwasaki A., Stiernholm B. J., Chan A. K., et al. Enhanced CTL responses mediated by p lasmid DNA immunogens encoding co-stimulatory molecules and cytokines. J Immuno l, 1997,158 (10) : 4591-4601.
[86] Simon J. H.,Nall B. C., Sony T. A.,et al. Direct Exposure of Therapy Mouse Spermatogenic cells to High Doses of Adenovirus Gene. J.Immuno l, 2000, 164 (6) : 3102-3111. Human, et al.
[87] Anim Kevin R. S. Sperm cell mediated transgenesis : a review., Biotechnology1999 , (10) :1～13.
[88]高华颖,曹阳,李世辉等.精子介导转基因动物的制备.遗传.2003,25 (3) :283～290.
[89] Izsvak Z Ivics Z. Sleeping Beauty transposition: biology and applications for molecular therapy. Mol Ther, 2004, 9(2): 147-156.
[90]Adam J.D.,Karl C.,Corey M.,et al.Mammalian germ-line transgenisis by transpostion.2002,99(7):4495-4499.
[91] Tang D., David M., Johnson SL.Genetic immunization is simple method for eliciting an immune response. Nature, 1992, 356: 152-154.
[92] Hinrichs J.,Berger S.,Shaw J.G. Induction of antibodies to plant viral proteins by DNA-based immunization. 1997 J Virol. Methods, 66(2): 195-202.
[93] Yeung S.J., Anderson, J., Kobayashi, K., Oka, K. and Chan, L. Production of rabbit polyclonal antibody against apobec-1 by genetic immunization. J Lipid Res. 1997;38(5):2627-2632.
[94] Sundaram P., Xiao, W., Brandsma, J. Particle-mediated delivery of recombinant expression vectors to rabbit skin induces high-titered polyclonal antisera (and circumvents purification of a protein immunogen). Nucleic Acids Res ,1996,24(7), 1375-1377.
[95] Boussif.O., Lezoualch.F., Zanta M.A. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo:polyethylenimine.Proc Natl Acad Science USA,1995,92(16):7297-7301.
[96]谢飞,高波,宋成义等.“睡美人”转座子的研究进展.遗传,2007,29(7):785-792.
[97] Keita M. Seiji Y., Masayuki E.Transposon-independent increase of transcription by the Sleeping Beauty transposase.Biochem Biophys Res Commun 2004,317(3):796–800.
[98]赵武,肖少波,方六荣等.牛疱疹病毒Ⅰ型VP22和猪繁殖与呼吸综合征病毒GP5融合基因DNA疫苗的免疫效应.生物工程学报,2005,21(5):725-730.
[99] Gary T. B., Carlota D., Martha S. Extended immunization intervals enhance the immunogenicity and protective efficacy of plasmid DNA vaccines. Microbes and Infection.2007,29 (9)1439-1446.
[2] Yant S. R., Meuse L., Chiu W., et al. Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system. Nat Genet, 2000, 25(1): 35-41.
[3] Fischer S. E., Wienholds E., Plasterk R. H. Regulated transposition of a fish transposon in the mouse germ line. Proc Natl Acad Sci USA, 2001,98(12): 6759-6764.
[4] Sheng D., Hui W. X., Li G., et al. Efficient transposition of the piggyBac resource (PB) transposon in mammalian cells and mice. Cell, 2005, 122(3): 473-483.
[5] Ivics Z., Hackett P. B., Plasterk R. H.,et al. Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Cell, 1997, 91(4): 501-510.
[6] Horie K., Yusa K., Yae K., et al. Characterization of Sleeping Beauty transposition and its application to genetic screening in mice. Mol Cell Biol, 2003, 23(24): 9189-9207.
[7] Izsvak Z., Ivics Z,, Plasterk R. H. Sleeping Beauty, a wide host-range transposon vector for genetic transformation in vertebrates. Mol Biol, 2000, 302: 93-102.
[8] Liu L., Mah C., Fletcher B. S. Sustained FVIII expression and phenotypic correction of Hemophilia A in neonatal mice using an endothelial-targeted Sleeping Beauty transposon. Mol Ther, 2006, 13(5): 1006-1015.
[9]丁昇,吴晓晖.“睡美人”转座系统研究进展.生物化学与生物物理进展,2003,30(1):43-48.
[10] Baus J., Liu L., Arnold D., et al.Hyperactive transposase mutants of the Sleeping Beauty Transposon. Mol Ther, 2005, 12(6): 1148-1156.
[11] Cui Z., Geurts A. M., Liu G., et al.Structure-function analysis of the inverted terminal repeats of the sleeping beauty transposon. Mol Biol, 2002, 318(5): 1221-1235.
[12] Geurts A. M., Yang Y., Clark K. J., et al. Gene transfer into genomes of human cells by the Sleeping Beauty transposon system. Mol Ther, 2003, 8(1): 108-117.
[13] Zayed H., Izsvak Z., Khare D., et al.The DNA bending protein HMGB1 is a cellular cofactor of Sleeping Beauty transposition. Nucleic Acids Res, 2003, 31(9): 2313-2322.
[14] Dupuy A. J., Fritz S., Largaespada D. A. Transposition and gene disruption in the male germline of the mouse. Genesis, 2001, 30(2): 82-88.
[15] Horie K., Kuroiwa A., Ikawa M., et al. Efficient chromosomal transposition of a Tc1/mariner- like transposon Sleeping Beauty in mice. Proc Natl Acad Sci USA, 2001, 98(16): 9191-9196.
[16] Doak T. G., Doerder F. P., Jahn C. L., et al. A proposed superfamily of transposase genes:transposon-like element in ciliated protozoa and a common“D35E”motif. Proc Natl Acad Sci USA, 1994, 91(3): 942-946.
[17] Masuda K., Yamamoto S., Endoh M., et al. Transposon independent increase of transcription by the Sleeping Beauty transposase.Biochem. Biophys Res Commun, 2004, 317(3): 796-800.
[18] Vigdal T. J., Kaufman C. D., Izsvak Z., et al.Common physical properties of DNA affecting target site selection of Sleeping Beauty and other Tc1/mariner transposable elements. Mol Biol, 2002, 323(3): 441-452.
[19] Izsvak Z., Khare D., Behlke J., et al. Involvement of a bifunctional, paired-like DNA-binding domain and a transpositional enhancer in Sleeping Beauty transposition. Biol Chem, 2002, 277(37): 34581-34588.
[20] Liu L., Sanz S., Heggestad A. D., et al.Endothelial targeting of the Sleeping Beauty transposon within lung. Mol Ther, 2004, 10(1): 97-105.
[21] Izsvak Z., Stuwe E. E., Fiedler D., et al.Healing the wounds inflicted by Sleeping Beauty transposition by double-strandbreak repair in mammalian somatic cells. Mol Cell, 2004, 13(2): 279-290.
[22] Luo G., Ivics Z., Izsvak Z., et al. Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. Proc Natl Acad Sci USA, 1998, 95(18): 10769-10773.
[23] Liu G., Cui Z., Aronovich E. L., et al.Excision of Sleeping Beauty transposons: parameters and applications to gene therapy. Gene Med, 2004, 6(5): 574-583.
[24] Yant S. R., Wu X., Huang Y., et al. High-resolution genome-wide mapping of transposon integration in mammals. Mol Cell Biol, 2005, 25(6): 2085-2094.
[25] Geurts A. M., Hackett C. S., Bell J. B., et al. Structure-based prediction of integration-site preferences of transposons into chromosomes.Nucleic Acids Res, 2006, 34(9): 2803-2811.
[26] Miller D. G., Rutledge E. A., Russell D. W. Chromosomal effects of adeno-associated virus vector integration. Nat Genet, 2002, 30(22): 147-148.
[27]肖巧学,曹阳,黄自然,刘良式.昆虫中的mariner类转座子.遗传, 2000, 22(2): 114-118.
[28] Wilson M. H., Kaminski J. M., George A. L. "Functional zinc finger/sleeping beauty transposase chimeras exhibit attenuated overproduction inhibition." FEBS Lett, 2005, 579(27): 6205-6209.
[29] Yusa K Takeda J, Horie K. Enhancement of Sleeping Beauty transposition by CpG methylation: possible role of heterochromatin formation. Mol Cell Biol, 2004, 24(9): 4004-4018.
[30] Park C. W., Park J., Kren B. T., et al. Sleeping Beauty transposition in the mouse genome is associated with changes in DNA methylation at the site of insertion. Genomics, 2006, 88(2): 204-213.
[31] Hacein B. A., Svon K. C., Schmidt M., et al. LMO2-associated clonal T cellproliferation in two patients after gene therapy for SCID-X1. Science, 2003, 302(5644): 415-419.
[32] Dupuy A .J., Clark K., Carlson C. M., et al.Mammalian germ-line transgenesis by transposition. Proc Natl Acad Sci USA, 2002, 99(7): 4495-4499.
[33] Wilber A. C., Frandsen J. L., Geurts J. L, et al. RNA as a source of transposase for Sleeping Beauty mediated gene insertion and expression in somatic cells and tissues. Mol Ther, 2006, 13(3): 625-630.
[34] Score P. R., Belur L. R., Frandsen J. L., et al. Sleeping Beautymediated transposition and long-term expression in vivo: use of the Cre-LoxP recombinase system to distinguish transposition-specific expression. Mol Ther, 2006, 13(3): 617-624.
[35] Chen Z. J., Kren B. T., Wong P. P., et al. Sleeping Beauty-mediated down-regulation of huntingtin expression by RNA interference. Biochem Biophys Res Commun, 2005, 329(2): 646-652.
[36] Yant S. R., Ehrhardt A., Mikkelsen J. G., et al. Transposition from a gutless adeno-transposon vector stabilizes transgene expression in vivo. Nat Biotech, 2002, 20(10): 999-1005.
[37] Yae K., Keng V. W., Koike M., et al. Sleeping Beauty transposon-based phenotypic analysis of mice: lack of arpc3 results indefective trophoblast outgrowth. Mol Cell Biol, 2006, 26(16): 6185-6196.
[38] Dupuy A . J., Jenkins N. A., Copeland N. G. Sleeping Beauty: a novel cancer gene discovery tool. Hum Mol Genet, 2006, 15(Suppl. 1): R75-R79.
[39] Gordon J. W., Scangos G. A ., P lotkin D. J., et al. Genetic transformation ofmouse embryo by m icro injection of purified DNA. P roc Natl Acad Sci U SA , 1980, 77: 7280- 7384.
[40] Palmiter R. D., Brinster R. E., HammerM. E., et al. D ramatic growth of m ice that developed from eggs micronjected with metallothionein-grow th hormone fusiongenes. Nature, 1982, 300: 611- 615.
[41]庞有志,赵淑娟.转基因技术与鸡的抗病育种.中国兽医科技, 1999, 3: 43- 44.
[42]桑润滋主编.动物繁殖生物技术.北京:中国农业出版社2002.
[43] Chan A. W. S., Homan E. J., Ballou L. U., et al. Transgenic cattle p roduced by reverse-transcribed gene transfer in oocytes. Proc Natl Acad Sci U SA , 1998, 95: 14082 -14033.
[44] Chan A. W. S., Chong K. Y., M artinovich C., et al. T ransgenic monkey p roduced by retroviral gene transfer into mature oocytes. Science, 2001, 291: 309- 312.
[45]成勇,成国祥,张靖溥等.牛As1-酪蛋白-乙肝病毒表面抗原融和基因在转基因羊中的表达.生物工程学报, 1997,13 (2) : 154-159.
[46]崔保安,王莉娟,赵楠等.转基因动物关键技术及其进展.河南畜牧兽医, 2003, (4) : 10-11.
[47]严泉剑.动物转基因与核移值技术的新进展.动物医学进展2002, 23 (3) : 24-29.
[48]杨章平,常洪.转基因动物的研究进展.世界农业, 2001, (3) :39-42.
[49] Janeisch R., Mintz B. Simian virus 40 DNA sequences in DNA healthy adultm ice derived from preimplantation blastocysts injected with viral DNA. P roc N atl A cad Sci U SA , 1974, 71 (4) :1250.
[50]刘谦,朱鑫泉.生物安全.北京:科学出版社, 2001.
[51] Schnieke A. E., Kind A. J., R itchie W. A ., et al. Human facto r IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblaatsp. Science, 1997, 278: 2130-2133.
[52]沈子龙,廖建民,徐寒梅.转基因动物技术与转基因动物制药.中国药科大学学报, 2002, 33 (2) : 81-86.
[53] Brakette B. G., Braransca W., Sawicki W. Uptake of heterologous genome by mammalian spermatozoa and its transfer to ova through fertilization. Proc. Natl. Acad. Sci. USA. ,1971 ,68 (2) :353-357.
[54] Arezzo F. Sea urchin sperm as a vector of foreign genetic information. Cell.Biol. Int . Rep. 1989 , 13 (4) :391 - 404.
[55] Lavitrano M., Camaioni A., Fazio V. M., et al . Sperm cells as vectors for introducing foreign DNA into eggs. Genetic transformation of mice. Cell , 1989 ,57 :717-723.
[56] Brackett B. G., Baranska W., Sawicki W., et al. Up take of hetero logous genome by mammallan spermatozoa and its transfer to ova through fertlization. P roc N atl A cad SciU SA , 1971, 68: 353-357.
[57] Anthony C. F., Teruhiko W. Mammali transgenesis by intracyto plasmic sperm injection. Science, 1999, 284: 1180-1183.
[58] Kim J. H., Jung H. H., Lee H. T. Development of a posi-mousetive method for male stem cell-mediated gene transfer in pig.Mol Reprod Dev.1997 , 46 (4) :515-526.
[59] Yamaza K.Y., Fujimoto H., Ando H. In vivo gene, acid transfer to mouse spermatogenic cells by deoxyribonucleic, acid injection into seminiferous tubules and subsequent elect roporation . Biol Reprod ,1998 ,59 (6) :1439-21444.
[60]沈新民,乔贵林,张玲.用曲细精管微注射法建立绿色荧光蛋白,转基因小鼠.第一军医大学学报.2002 , 22 (3) :250-2253.
[61] Garrett F. E., Goel S., Yasu l. J., et al . Liposomes fuse, impermeant with sperm cells s and induce activation by delivery of agents. Biochim Biophys Acta 1999 , 72 (3) :435-444.
[62]肖红卫,郑新民,陈思怀等.精子介导生产转基因猪.华中农业大学学报.2006,25(2):170-173.
[63] HE X., QI B., LIU G. S., YU W. D., et al. A Novel Method to , Transfer Gene In vivo System. Progress in Biochemistry and Biophysics 2006, 33(7): 685-690.
[64] Atkinson P. W., Hines E. R., Beaton S., et al. Association of exoigenous DNA with cattle and insect spermatozoa in vitro. Mo l Rep rod , 1991, 29 (1) : 1-5.
[65] Lavitrano M.,French D., Zani M., et al. The interaction between exogenousDNAand sperm cells. Mo l Reprod, 1992, 31: 161-169.
[66]翟少伟,谷艳钗.转基因动物的研究进展和前景展望.当代畜牧, 2000, (6) : 123.
[67] Arezzo F. Sea urchin sperm as a vector of foreign genetic informatiom . Cell Biol Int Reprod 1989 , 13 :391 - 404.
[68] Nakanishi A., IritaniA. Gene reansfer in the ch icken by sperm2mediated methods. Mo l Repo rd, 1993,36 (2) : 258-261.
[69] Ebert K. M., Selgrath J. P. Transgenic production of a variant of human tissue-type plasm inogen activator in goat milk: generation of transgenic goats and analysis of expression. Bio techno logy,1991, (9) : 835-838.
[70] Bach iller D., Schellander K., Peli J., et al. L iposome Mediated DNA up take by sperm. Mo l Rep rod,1991, 30 (3) : 194-200.
[71] Wolff J. A., Malone R. W., Williams P., et al. Direct gene transfer into mouse muscle in vivo. Science ,1990 ,247∶1465 -1468.
[72] Hartikka J., Sawdey M., Jensen F. C.,et al. An improved plasmid DNA expression vector for direct inection into skeletal muscle. HumanGene Therapy ,1996 ,7∶1205 -1209.
[73] Benvenisty B., Reshef. Direct introduction of genes into rats and expression of the genes. P roc Natl Sci USA. 1986 12, 83 (24) : 9551-9555.
[74] Guyla A., George D., Donald R. Human dystroph in expression in mdx mice after intramuscular injection of DNA constructs. Natrue, 1991, 352: 815-818.
[75] Tang C., Devit M., Johnston S. A. Genetic immunization is a simple method for eliciting an immune response. Nature,1992, 356 (6365) : 152-154.
[76] Ulmer J. B., Donnelly J. J., Parker S. E., et al. Hetero logous protection against influenza by injection of DNA encoding a viral protein. Science, 1993, 259 (5102) : 1745-1749.
[77]王世若.当前研制预防用疫苗的发展趋势.预防兽医学进展2000 , 30(3)∶15.
[78]解洪业.新型疫苗的研究与发展.青海畜牧兽医杂志1997 , (3)∶24—26.
[79] Schrijver R., Langendijk J. P., Keil G.M, et al. Immunization ofcattle with a BHVi vector vaccine or a DNA vaccine both coding for the G-protein of BR.Vaccine ,1997 ,15∶1908-1916.
[80] Krieg A.M., YiA K., Matson S., et al. CpG motifs in bacterial DNA trigger direct B cell activation. Nature, 1995, 374(6522) : 546-569.
[81] Temperton N. J., Quenelle D.C., Lawson K.M., et al. Enhancement of humoral immune responses to a human cytome galovirus DNA vaccine: Adjuvant effects of aluminum phosphate and CpG oligodeoxynucleo tides. J Med V iro l,2003, 70 (1) : 86-90.
[82] Youssef S., Wildbaum G., Maor G., et al. Long lasting protective immunity to experimental auto immune encephalo myelitis following vaccination with naked DNA encoding C-C chemokines. J Immunol, 1998, 161 (8) : 3870-3879. .
[83] Okada E. SasakiS, Ish ii N., et al. Intranasal immunization of a DNA vaccine with IL -12 and granulocyte macrophage colony stimulating factor (GM-CSF) expressing plasmids in liposomes against HIV-1 antigens. J. Immunol, 1997, 159 (70) : 3638-3647.
[84] Kusakabe K., Xin K. Q., Katoh H., et al. The timing of GM-CSF expression plasmid administration influences the Th1?T h rsponse induced by an H IV -1 specific DNA vaccine. J.Immuno l, 2000, 164 (6) : 3102-3111.
[85] Iwasaki A., Stiernholm B. J., Chan A. K., et al. Enhanced CTL responses mediated by p lasmid DNA immunogens encoding co-stimulatory molecules and cytokines. J Immuno l, 1997,158 (10) : 4591-4601.
[86] Simon J. H.,Nall B. C., Sony T. A.,et al. Direct Exposure of Therapy Mouse Spermatogenic cells to High Doses of Adenovirus Gene. J.Immuno l, 2000, 164 (6) : 3102-3111. Human, et al.
[87] Anim Kevin R. S. Sperm cell mediated transgenesis : a review., Biotechnology1999 , (10) :1～13.
[88]高华颖,曹阳,李世辉等.精子介导转基因动物的制备.遗传.2003,25 (3) :283～290.
[89] Izsvak Z Ivics Z. Sleeping Beauty transposition: biology and applications for molecular therapy. Mol Ther, 2004, 9(2): 147-156.
[90]Adam J.D.,Karl C.,Corey M.,et al.Mammalian germ-line transgenisis by transpostion.2002,99(7):4495-4499.
[91] Tang D., David M., Johnson SL.Genetic immunization is simple method for eliciting an immune response. Nature, 1992, 356: 152-154.
[92] Hinrichs J.,Berger S.,Shaw J.G. Induction of antibodies to plant viral proteins by DNA-based immunization. 1997 J Virol. Methods, 66(2): 195-202.
[93] Yeung S.J., Anderson, J., Kobayashi, K., Oka, K. and Chan, L. Production of rabbit polyclonal antibody against apobec-1 by genetic immunization. J Lipid Res. 1997;38(5):2627-2632.
[94] Sundaram P., Xiao, W., Brandsma, J. Particle-mediated delivery of recombinant expression vectors to rabbit skin induces high-titered polyclonal antisera (and circumvents purification of a protein immunogen). Nucleic Acids Res ,1996,24(7), 1375-1377.
[95] Boussif.O., Lezoualch.F., Zanta M.A. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo:polyethylenimine.Proc Natl Acad Science USA,1995,92(16):7297-7301.
[96]谢飞,高波,宋成义等.“睡美人”转座子的研究进展.遗传,2007,29(7):785-792.
[97] Keita M. Seiji Y., Masayuki E.Transposon-independent increase of transcription by the Sleeping Beauty transposase.Biochem Biophys Res Commun 2004,317(3):796–800.
[98]赵武,肖少波,方六荣等.牛疱疹病毒Ⅰ型VP22和猪繁殖与呼吸综合征病毒GP5融合基因DNA疫苗的免疫效应.生物工程学报,2005,21(5):725-730.
[99] Gary T. B., Carlota D., Martha S. Extended immunization intervals enhance the immunogenicity and protective efficacy of plasmid DNA vaccines. Microbes and Infection.2007,29 (9)1439-1446.
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