人类卵母细胞报告系统的构建

doi:10.13560/j.cnki.biotech.bull.1985.2015.11.033

生物技术通报 ›› 2015, Vol. 31 ›› Issue (11): 251-256.doi: 10.13560/j.cnki.biotech.bull.1985.2015.11.033

人类卵母细胞报告系统的构建

罗梦圆, 纪家葵

清华大学医学院干细胞与再生医学中心，北京 100084

收稿日期:2015-03-25 出版日期:2015-11-26 发布日期:2015-11-26
作者简介:罗梦圆，女，硕士研究生，研究方向: 干细胞与生殖发育；E-mail: lqqsqslkmzcz@mail.bnu.edu.cn

Constructing a Human Oocyte Reporter System

Luo Mengyuan, Kee Kehkooi

Center for Stem Cell Biology and Regenerative Medicine，School of Medicine，Tsinghua University，Beijing 100084

Received:2015-03-25 Published:2015-11-26 Online:2015-11-26

摘要/Abstract

摘要： 体外分化培养干细胞得到成熟生殖细胞具有重要的科学价值和临床意义，构建出人类胚胎干细胞体外分化为卵细胞的报告系统，旨在能为这一研究提供有效的工具。ZP2是组成哺乳动物卵细胞周围透明带的重要蛋白之一，其蛋白只能在卵细胞中表达，具有很高的组织特异性。将人ZP2基因300 bp和2 500 bp启动子片段分别与eGFP荧光蛋白基因连接，利用Gateway 系统构建出重组质粒ZP2-300-eGFP和ZP2-2500-eGFP，在WI38、未分化的人胚胎干细胞H9和小鼠卵母细胞中验证其荧光蛋白表达组织特异性。结果表明，ZP2-2 500-eGFP报告系统可以特异启动荧光报告基因在小鼠的卵母细胞中的表达。

关键词: 报告系统, ZP2, 卵母细胞

Abstract: In vitro differentiation of stem cells into mature germ cells has important significance both in scientific research and in clinical treatment. An oocyte reporter system will become a useful tool for this goal. ZP2 is a component of the zona pellucida in mammalian oocytes and its expression is known to be restricted in oocyte. In this experiment we cloned 300 bp and 2 500 bp fragment of human ZP2 promoter and fused them to eGFP aiming to build an oocyte reporter sysem. We tested the specificity of the reporter in WI38, undifferentiated human embryonic stem cell(hESC)line H9 and mouse oocyte. Results show that ZP2-2500-eGFP can specifically drive eGFP expression in mouse oocyte but no eGFP was detected in WI38 and undifferentiated hESCs.

Key words: reporter system, ZP2, oocyte

罗梦圆, 纪家葵. 人类卵母细胞报告系统的构建[J]. 生物技术通报, 2015, 31(11): 251-256.

Luo Mengyuan, Kee Kehkooi. Constructing a Human Oocyte Reporter System[J]. Biotechnology Bulletin, 2015, 31(11): 251-256.

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人类卵母细胞报告系统的构建

Constructing a Human Oocyte Reporter System

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