遗传 >

2009 , Vol. 31 >Issue 6: 595 - 599

DOI: https://doi.org/10.3724/SP.J.1005.2009.00595

研究报告

卵母细胞特异表达fC31整合酶载体pZP3-INT的构建及其在小鼠卵母细胞中的表达

展开
  • 上海交通大学医学遗传研究所, 卫生部医学胚胎分子生物学重点实验室, 上海市胚胎与生殖工程重点实验室, 上海200040

收稿日期: 2009-03-19

  修回日期: 2009-04-10

  网络出版日期: 2009-06-10

收起

Construction of the oocyte-specific expressing fC31 integrase vector pZP3-INT and its expression in mouse oocytes

Expand
  • Shanghai Institute of Medical Genetics, Shanghai JiaoTong University, Key Laboratory of Embryo Molecular Biology, Ministry of Health, Shanghai Laboratory of Embryo and Reproduction Engineering, Shanghai 200040, China

Received date: 2009-03-19

  Revised date: 2009-04-10

  Online published: 2009-06-10

Fold

摘要

链霉菌噬菌体fC31整合酶是一种位点特异性重组酶(Site-specific recombinase, SSR), 可介导链霉菌噬菌体attP位点(Phage attachment site)和链霉菌基因组attB位点(Bacterial attachment site)间的单向重组。为探讨它能否应用于卵母细胞特定基因的重组, 文章采用卵巢针刺取卵法采集生发泡(GV)期小鼠卵母细胞, 将卵透明带糖蛋白3(ZP3)启动子驱动的fC31整合酶表达载体pZP3-INT和检测fC31整合酶位点特异性重组功能的重组质粒载体pBCPB+, 通过显微注射导入到小鼠卵母细胞中。培养48 h后, RT-PCR检测fC31整合酶mRNA表达以及PCR检测pBCPB+载体发生重组的情况。结果表明: 载体pZP3-INT在卵母细胞中表达fC31 整合酶mRNA; 并且pBCPB+载体发生了位点特异性重组, 提示fC31整合酶在卵母细胞中可以介导位点特异性重组反应。

关键词： ΦC31整合酶; 位点特异性重组; 基因操作; 卵透明带糖蛋白3; 卵母细胞

本文引用格式

徐焕宇，龚秀丽，郭歆冰，马晴雯，曾溢滔 . 卵母细胞特异表达fC31整合酶载体pZP3-INT的构建及其在小鼠卵母细胞中的表达[J]. 遗传, 2009 , 31(6) : 595 -599 . DOI: 10.3724/SP.J.1005.2009.00595

Abstract

Streptomyces phage ΦC31 integrase is a site-specific recombinase, which can catalyze site-specific, unidirectional recombination between the attP site and attB site. To explore whether it can be used to mediate the recombination of specific gene in oocytes, GV-stage oocytes were collected from 3-week-old Kunming White mice by puncturing antral follocles with a sharp needle, and micro-injected with oocyte-specific expressing ΦC31 integrase vector pZP3-INT and site -specific recombination detection vector pBCPB+. ΦC31 integrase mRNA were detected by RT-PCR and the recombination of pBCPB+ was evaluated by PCR in mouse oocytes at 48 h after injection. Both can get corresponding bands. These results indicated that the expression of ΦC31 integrase can be driven by ZP3 promoter efficiently and ΦC31 integrase can mediate the site-specific recombination between attP site and attB site in mouse GV-stage oocytes. It could be a powerful tool for the study of recombination of specific gene in mouse oocytes and would provide an alternative way for the mouse oocyte genome manipulation.
Options
文章导航

/

〈 〉

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

版权所有 © 中国科学院遗传与发育生物学研究所 中国遗传学会
地址：北京市朝阳区北辰西路1号院　中国科学院遗传与发育生物学研究所1号楼　
邮政编码：100101
联系电话：010-64807669
E-mail：yczz@genetics.ac.cn