牛结蛋白基因启动子的初步改造研究  

The Study of Recombination of Bovine Desmin Gene Promoter

作　　者：亓利思 佟慧丽[1] 李树峰[1] 胡倩[1] 严云勤[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《畜牧兽医学报》2016年第12期2405-2413,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：基金项目:高产优质转基因肉牛新品种培育(2014ZX08007-002)

摘　　要：本研究在前期研究工作的基础上,通过添加特异的调控元件对牛结蛋白(Desimn)基因启动子进行改造,以期获得活性较高且具有肌肉组织特异性的启动子。本研究以300bp的牛结蛋白基因启动子为基础,对其进行改造,即通过串联多拷贝的正调控元件的方式,成功构建了3个改造后的结蛋白基因启动子片段:pGL3-desmin300-desmin180、pGL3-MD-αD1-desmin300和pGL3-αD2-αD1-desmin300。结果表明,pGL3-desmin300-desmin180、pGL3-MD-αD1-desmin300和pGL3-αD2-αD1-desmin300的启动子活性分别是野生型启动子pGL3-desmin300的1.67、2.56和2.88倍,说明改造后的启动子片段具有较高的启动活性。同时,牛骨胳肌卫星细胞和成纤维细胞试验结果表明,以上3组改造后的启动子片段具有较强的肌肉特异性,以上高效肌肉特异性启动子片段的应用为提高肌肉产量的转基因家畜肉的生产提供重要帮助。On the basis of previous results,we transformed a bovine Desminpromoter fragment to obtain some high efficiency and muscle tissue-specific promoters which could be useful.We obtained 3artificial Desminpromoter fragments successfully through adding several positive specific regulatory elements into the bovine Desmin promoter of 300 bp length,which included pGL3-desmin300-desmin180,pGL3-MD-αD1-desmin300 and pGL3-αD2-αD1-desmin300.The results showed that the activities of pGL3-desmin300-desmin180,pGL3-MD-αD1-desmin300 and pGL3-αD2-αD1-desmin300 were 1.67,2.56 and 2.88 times more than activity of pGL3-desmin300,respectively.Bovine skeletal muscle satellite cells and fibroblast cell experiments showed that the3 artificial Desmin promoter fragments had high muscle specificity.The application of Desmin fragments with high efficiency and muscle specificity will provide an important help to increase the output of muscle production of transgenic livestock.

关 键 词：牛 结蛋白 调控元件 肌肉特异性 双荧光报告检测 

分 类 号：S823[农业科学—畜牧学] S813.2[农业科学—畜牧兽医]