牛结蛋白基因启动子的初步改造研究

doi:10.11843/j.issn.0366-6964.2016.12.010

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (12): 2405-2413.doi: 10.11843/j.issn.0366-6964.2016.12.010

牛结蛋白基因启动子的初步改造研究

亓利思，佟慧丽，李树峰,胡倩，严云勤^*

（东北农业大学生命科学学院，哈尔滨 150030）

收稿日期:2016-03-22 出版日期:2016-12-23 发布日期:2016-12-23
通讯作者: 严云勤，教授，博士生导师，主要从事动物转基因的研究，E-mail:yanyunqin@sohu.com
作者简介:亓利思(1991-)，男，山东莱芜人，硕士，主要从事分子克隆与启动子活性的研究，E-mail:1027916478@qq.com
基金资助:
高产优质转基因肉牛新品种培育(2014ZX08007-002)

The Study of Recombination of Bovine Desmin Gene Promoter

QI Li-si, TONG Hui-li, LI Shu-feng, HU Qian, YAN Yun-qin^*

(College of Life Science, Northeast Agricultural University, Harbin 150030,China)

Received:2016-03-22 Online:2016-12-23 Published:2016-12-23

摘要/Abstract

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300 bp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。

Abstract:

On the basis of previous results, we transformed a bovine Desmin promoter fragment to obtain some high efficiency and muscle tissue-specific promoters which could be useful. We obtained 3 artificial Desmin promoter 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 the 3 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.

中图分类号:

S823
S813.2

亓利思，佟慧丽，李树峰,胡倩，严云勤. 牛结蛋白基因启动子的初步改造研究[J]. 畜牧兽医学报, 2016, 47(12): 2405-2413.

QI Li-si, TONG Hui-li, LI Shu-feng, HU Qian, YAN Yun-qin. The Study of Recombination of Bovine Desmin Gene Promoter[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(12): 2405-2413.

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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

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