myostatin基因打靶的成肌细胞制备

中国生物工程杂志

2009, Vol. 29

Issue (07): 22-26

研究报告

myostatin基因打靶的成肌细胞制备

张蕾1|杨兴元2|安晓荣2|陈永福2

1. 辽宁医学院 2. 中国农业大学生物学院农业生物技术国家重点实验室

The Construct of Myostatin Gene Targeted Recipient Myoblast Cell

ZHANG Lei1|YANG Xing-yuan2|AN Xiao-rong2|CHEN Yong-fu2

1. Liaoning Medical University,Jinzhou121000,China 2. State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University,Beijing100094,China

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摘要： Myostatin(MSTN), β-转化生长因子超家族的一员，是肌肉生长的负调控因子，该基因自然突变的比利时蓝牛和皮尔蒙特牛出现双肌性状。基因敲除该基因，是实现家畜产肉量的提高的有效方法。通过建立无启动子打靶载体MSTN-GFP, MSTN-neo，转染新生和胎儿绵羊骨骼肌细胞，经过表达绿色荧光蛋白报告基因和G418筛选获得骨骼肌细胞克隆，PCR，Southern blot,DNA序列检测获得阳性细胞克隆，为制备myostatin基因敲除的体细胞克隆绵羊提供核移植供体。

关键词： 基因打靶;myostatin;成肌细胞

Abstract: Abstract Myostatin (MSTN), a member of the transforming growth factor-β（TGF-β）superfamily, has been shown to be a negative regulator of myogenesis. Natural mutation in beef cattle causes double-muscling phenotypes. An investigation designed to knock out the myostatin gene by gene targeting in ovine myoblast cells were reported. Two promoter-trap targeting vectors MSTN-GFP and MSTN-Neo were constructed and used to transfect fetal and neonatal ovine primary myoblast cells. Both GFP expressing cells and drug resistant cells were obtained. Targeted cells expressing GFP were confirmed by PCR assay and drug resistant cells were characterized by PCR, Southern blot and DNA sequnce detection after growing into cell clones. That would supply nuleus transplantation donator for myostatin gene targetted somatic cloning sheep.

Key words: Gene targeting Myostatin Myoblast cell

收稿日期: 2008-12-15 出版日期: 2009-07-28

ZTFLH:

中图分类号Q813

基金资助: 国家“863”计划资助项目(2002AA206311)

通讯作者: 张蕾 E-mail: E-mail：zhbud@hotmail.com

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引用本文:

张蕾1,杨兴元2,安晓荣2,陈永福2. myostatin基因打靶的成肌细胞制备[J]. 中国生物工程杂志, 2009, 29(07): 22-26.

ZHANG Lei-1, YANG Xin-Yuan-2, AN Xiao-Rong-2, CHEN Yong-Fu-2. The Construct of Myostatin Gene Targeted Recipient Myoblast Cell. China Biotechnology, 2009, 29(07): 22-26.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2009/V29/I07/22

［1］ McCreath K J, Howcroft J, Campbell K H, et al.Production of gene-targeted shee Pby nuclear transfer from cultured somatic cells. Nature, 2000，405: 1066~1069 ［2］ Kambadur R, Sharma M, Smith T P,et al.Mutations in myostatin (GDF8) in Double-Muscled Belgian Blue and Piedmontese Cattle. Genome research,1997,7:910~915 ［3］ McPherron A C, Lawler A M, Lee S J.Regulation of skeletal muscle mass in mice by a new TGF-b superfamily member. Nature, 1997,387: 83~90 ［4］ Kaufman S J, Foster R F.Replicating myoblasts express a muscle-specific phenotype. PNAS,1998,85: 9606~9610 ［5］ Schwartzberg P L, Robertson E J, Goff S P. Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein. PNAS,1990,87:3210-3214 ［6］ Hanson K D, Sedivy J M. Analysis of biological selection for high- efficiency gene targeting. Mol Cell Biol,1995,15:45~51 ［7］ Denning C, Burl S, Ainslie A,et al.Deletion of the α-1,3 galactosyl transferase (GGTA1) gene and the prion protein(PrP) gene in sheep. Nature Biotech,2001,19: 559~562 ［8］ Jin D I, Lee S H, Choi J H,et al. Targeting efficiency of α-1,3-galactosyl transferase gene in pig fetal fibroblast cells. Experimental and Molecular Medicine,2003,35:572~577 ［9］ McFarland G A, Holliday R.Retardation of the senescence of cultured human diploid fibroblasts by carnosine. Exp Cell Res,1994, 212:167~175 ［10］ Morales C P, Holt S E. Ouellette M, et al. Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nat Genet,1999,21:115~118 ［11］ Bodnar A G, Ouellette M, Frolkis M, et al.Extension of life-span by introduction of telomerase into normal human cells.Science,1998,279: 349~352 ［12］ Wyllie F S, Jones C J, Skinner J W, et al.Telomerase prevents the accelerated cell ageing of Werner syndrome fibroblasts. Nat Genet,2000,24: 16~17

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