›› 2013, Vol. 40 ›› Issue (9): 183-187.

• 遗传繁育 • 上一篇    下一篇

人α-乳清蛋白转基因克隆牛外源基因拷贝数的分析

王洁雯1, 王敬姣1, 赵姝君1, 李宁2, 李世杰1, 张萃1   

  1. 1. 河北农业大学生命科学学院, 河北保定 071001;
    2. 中国农业大学农业生物技术国家重点实验室, 北京 100193
  • 收稿日期:2013-02-27 出版日期:2013-09-20 发布日期:2013-09-18
  • 通讯作者: 张萃 E-mail:zhangcui@hebau.edu.cn
  • 作者简介:王洁雯(1987-),女,河北人,硕士生,研究方向:基因工程。
  • 基金资助:
    河北省自然基金项目(C2011204001);转基因生物新品种培育重大专项(2011ZX08009-002、2009ZX0801024B)。

Detecting Copy Number of Foreign Gene in Human Alpha-lactalbumin Transgenic Cloned Cattle

WANG Jie-wen1, WANG Jing-jiao1, ZHAO Shu-jun1, LI Ning2, LI Shi-jie1, ZHANG Cui1   

  1. 1. College of Life Science, Hebei Agricultural University, Baoding 071001, China;
    2. State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China
  • Received:2013-02-27 Online:2013-09-20 Published:2013-09-18

摘要: 为了探讨转基因克隆动物外源基因拷贝数及拷贝数在传代过程中的变化,本研究应用实时荧光定量PCR检测人α-乳清蛋白(human alpha-lactalbumin,HLA)转基因克隆牛及其子代共5头牛中外源基因的拷贝数,其中K060923为亲代,172、081、189、隆娃为子代。HLA基因在亲代转基因克隆牛K060923中的拷贝数为1.145,在子代转基因克隆牛172、081、189和隆娃中分别为0.998、1.107、0.891和0.842。结果表明,5头牛中外源基因拷贝数都在1左右,说明人α-乳清蛋白基本以单拷贝形式整合到宿主基因组中,且稳定遗传。

关键词: 人α-乳清蛋白基因; 转基因克隆牛; 实时荧光定量PCR; 拷贝数

Abstract: To explore the changes of the exogenous gene copy number in transgenic cloned animals during subculture, we detected the copy number of human alpha-lactalbumin gene in five transgenic cloned cattles, including a parental cattle(K060923) and four offspring cattles(172,081,189 and Longwa), with Real-time fluorescent quantitative PCR. Among the five cows, K060923 was parental and the rest are offspring. The copy number of K060923 was 1.145, and that of the four offspring (172, 081, 189 and Longwa) was 0.998, 1.107, 0.891 and 0.842, respectively. The obtained results indicated that the copy numbers of all the five cows were about one. The results showed that exogenous gene was integrated into the host genome as a single copy and was stably inherited.

Key words: human alpha-lactalbumin; transgenic cloned cattle; Real-time fluorescent quantitative PCR; copy numbe

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