高效率基因打靶载体的构建及受体成肌细胞的制备

J4 ›› 2009, Vol. 47 ›› Issue (6): 1328-1333.

高效率基因打靶载体的构建及受体成肌细胞的制备

张蕾¹, 杨兴元², 安晓荣², 陈永福²

1. 辽宁医学院基础医学院| 辽宁锦州 121000;2. 中国农业大学生物学院农业生物技术国家重点实验室, 北京 100094

收稿日期:2009-03-13 出版日期:2009-11-26 发布日期:2010-01-07
通讯作者: 张蕾 E-mail:zhbud@hotmail.com.

The Construct of Promotertrap and PolyAtrap Gene TargettingVector and the Preparation of Recipient Myoblast Cell

ZHANG Lei¹, YANG Xingyuan², AN Xiaorong², CHEN Yongfu²

1. Department of Basic Medical, Liaoning Medical University, Jinzhou 121000, Liaoning Province, China|2. State KeyLaboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100094, China

Received:2009-03-13 Online:2009-11-26 Published:2010-01-07
Contact: ZHANG Lei E-mail:zhbud@hotmail.com.

摘要/Abstract

摘要：

通过建立高效率“双无”（无启动子、无polyA）打靶载体MSTN-GFP和MSTNneo及新生绵羊和胎儿成肌细胞的培养和鉴定, 优化了培养条件, 改善了细胞状态, 为提高打靶效率及体细胞克隆提供了稳定的核移植供体；同时无启动子打靶载体的构建也有利于打靶后阳性细胞克隆的分子鉴定.

关键词: 无启动子打靶载体；肌肉抑制素；成肌细胞；基因打靶

Abstract:

We reported an investigation designed to knockout the MSTN gene by gene targeting in ovine myoblast cells. To increase the gene target efficiency and to supply nuleus transplantation donator for somatic cloning, two double traps (promotertrap and polyA trap) and targeting vectors MSTNgreen fluorescent protein (GFP) and MSTNneo were constructed and the foetal and neonatal ovine primary myoblast were cultured and identified. The cell cultural conditions were optimized. At the same time, the construct of no promoter vector was easy to molecular identification of positive cell clone after gene targetting.

Key words: promotertrap targeting vetor, Myostatin, myoblast cell, gene targenting

中图分类号:

Q813

张蕾, 杨兴元, 安晓荣, 陈永福. 高效率基因打靶载体的构建及受体成肌细胞的制备[J]. J4, 2009, 47(6): 1328-1333.

ZHANG Lei, YANG Xingyuan, AN Xiaorong, CHEN Yongfu. The Construct of Promotertrap and PolyAtrap Gene TargettingVector and the Preparation of Recipient Myoblast Cell[J]. J4, 2009, 47(6): 1328-1333.