牛β-酪蛋白座位无启动子基因打靶载体的构建

doi:10.7668/hbnxb.2010.04.004

华北农学报 ›› 2010, Vol. 25 ›› Issue (4): 17-24. doi: 10.7668/hbnxb.2010.04.004

所属专题： 畜牧；生物技术；

牛β-酪蛋白座位无启动子基因打靶载体的构建

李瑞国¹, 苗朝华², 侯健³, 关宏³, 安晓荣³, 陈永福³

1. 中国生物技术发展中心,北京 100036;
2. 中国农业科学院生物技术研究所,北京 100081;
3. 中国农业大学农业生物技术国家重点实验室,北京 100094

收稿日期:2010-07-02 出版日期:2010-08-28
通讯作者: 安晓荣(1957- )，女，山东济南人，博士，教授，主要从事动物生理学研究。
作者简介:李瑞国(1973- ),男,山西晋城人,副研究员,博士,主要从事动物生理学研究.
基金资助:
国家"863"计划项目(2002AA206311);国家自然科学基金项目(30871783;30700573)

Non-Promotor Gene Targeting Vector Construction for Bovine β-casein Site

LI Rui-guo¹, MIAO Chao-hua², HOU Jian³, GUAN Hong³, AN Xiao-rong³, CHEN Yong-fu³

1. China National Center for Biotechnology Development, Beijing 100036, China;
2. Biotechnology Research Institute, Chinese Academy of Agriculture Sciences, Beijing 100081, China;
3. State Key Lab for Agro-Biotechnology, China Agricultural University, Beijing 100094, China

Received:2010-07-02 Published:2010-08-28

摘要/Abstract

摘要： 将带有新霉素基因(neo)和绿色荧光蛋白基因(GFP)的质粒pEGFP-C1进行了改造,去掉neo基因的启动子(Psv40),在无启动子的neo和完整的GFP基因两侧分别加入一个LoxP位点.然后将带有LoxP位点、筛选标记基因和报告基因的结构装入左右分别为牛β-casein基因1 590 bp和5 33 bp的同源臂结构中,构建成无启动子基因打靶载体pT-1590-LoxP-GFP-LoxP-533.

关键词: 牛&beta, 酪蛋白, LoxP, 无启动子, 打靶载体, 构建

Abstract: Plasmid vector pEGFP-C1，including neo and GFP gene，was modified. Psv40 promoter for neo was deleted，and introduce a LoxP locus into each flank of non-promoter neo and complete GFP，constructed pEGFPLoxP. Then we introduced 1 590 bp and 5 833 bp sequences from bovine genomes into vector pEGFP-LoxP，completed construction of non-promoter gene targeting vector，which is pT-1590-LoxP-GFP-LoxP-5833，to bovine β-casein gene locus.

Key words: Bovine β-casein, LoxP, Non-promoter, Targeting vector, Construction

中图分类号:

Q785

李瑞国, 苗朝华, 侯健, 关宏, 安晓荣, 陈永福. 牛β-酪蛋白座位无启动子基因打靶载体的构建[J]. 华北农学报, 2010, 25(4): 17-24. doi: 10.7668/hbnxb.2010.04.004.

LI Rui-guo, MIAO Chao-hua, HOU Jian, GUAN Hong, AN Xiao-rong, CHEN Yong-fu. Non-Promotor Gene Targeting Vector Construction for Bovine β-casein Site[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(4): 17-24. doi: 10.7668/hbnxb.2010.04.004.

http://www.hbnxb.net/CN/Y2010/V25/I4/17

参考文献

[1] Rosen J M,Raught B,Hadsell D.The mammary gland as a bioreactor:factors regulating the efficient expression of milk protein based transgenes[J].Am J Clin Nutr(Supplement),1996,63:627-632.

[2] Sedivy J M,Vogelstein B,Liber H L,et al.Gene targeting in human cells without isogenic DNA[J].Science,1999,283:abstract.

[3] Sedivy J M,Sharp P A.Positive genetic selection for gene disruption in mammalian cells by homologous recombination[J].Pro Natl Acad Sci,USA,1989,86:227-231.

[4] Jeannotte L,Ruiz J C,Robertson E J.Low level of Hox1.3 gene expression does not preclude the use of promoterless vectors to generate a targeted gene disruption[J].Off Mol Cell Biol,1991,11:5578-5585.

[5] Shjichiri M,Hanson K D,Sedivy J M.Effects of c-myc expression on proliferation,qiuescence,and the G0 to G1 transition in nontransformed cells[J].Cell Growth Differ,1993,4:93-104.

[6] Maniatis T.Molecular Cloning:a Laboratory Manual(second edition)[M].New York:Cold Spring Harbor Laboratory Press,1989.

[7] 李瑞国,安晓荣,苟克勉,等.提高PCR产物T-A克隆效率的研究[J].农业生物技术学报,2003,11(2):158-162.

[8] McGreath K J,Howcroft J,campbell K H S,et al.Production of gene-targeted sheep by nuclear transfer from cultured somatic cells[J].Nature,2000,405:1066-1069.

[9] Denning C,Burl S,Ainslie A,et al.Deletion of the alpha(1,3) galactosyl transferase(GGTA1) gene and the prion protein(PrP) gene in sheep[J].Nat Biotechnol,2001,19:559-562.

[10] Te Riile H,Robanus M,Berns A.Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs[J].Pro Natl Acad Sci,USA,89:5128-5132.

[11] Deng C,Capecchi M R.Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus[J].Mol Cell Biol,1992,12:3365-3371.

[12] Yanez R J,Porter A C G.Therapeutic gene targeting[J].Gene Ther,1998,5:149-159.

[13] Shulman M J,Nissen L,Collins C.Homologous recombination in hybridoma cells:dependence on time and fragment length[J].Mol Cell Biol,1990,10:4466-4472.

[14] Thomas K R,Deng C,Capecchi M R.High-fedelity gene targeting in embryonic stem cells by using sequence replacement vectors[J].Mol Cell Biol,1992,12:2919-2923.

[15] Shjichiri M,Hanson K D,Sedivy J M.Effects of c-myc expression on proliferation,qiuescence,and the G0 to G1 transition in nontransformed cells[J].Cell Growth Differ,1993,4:93-104.

[16] Mansour S L,Thomas K R,Capecchi M R.Disruption of the protooncogene int-2 in mouse embryo-derived stem cells:a general strategy for targeting mutations to non-selectable genes[J].Natrue,1988,336:348-352.

[17] Thomas K R,Capecchi M R.Targeted disruption of the murine int-1 pro-oncogene resulting in severe abnormalities in midbrain and cerebellar development[J].Nature,1990,346:847-850.

[18] Hanson K D,Sedivy J M.Analysis of biological selections for high-efficiency gene targeting[J].Mol Cell Biol,1995,15:45-51.

[19] Revet B M,Sena E P,Zarling D A.Homologous DNA targeting with RecA protein coated short DNA probes and electronmicroscope mapping on linear duplex molecules[J].J Mol Biol,1993,232:779-791.

[20] Mekeel K L,Tang W,Kachnic L A,et al.Inactivation of p53 results in high rates of homologous recombination[J].Oncogene,1997,14:1847-1857.

[21] Xiang Y,Cole-Strauss A,Yoon K,et al.Targeted gene conversion in a mammalian CD34+-enriched cell population using a chimeric RNA/DNA oligonucleotide[J].J Mol Med,1997,75:829-835.

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