糖皮质激素受体M565R、C638S、L753F、A573Q、GR2439insA定点诱变体的构建及鉴定  

Construction and identification of site-directed mutants of glucocorticoid receptors M565R,C638S, L753F, A573Q and GR2349insA

作　　者：崔颖鹏[1] 陈培松[1] 欧阳涓[1] 黄彬[1] 周青松[1] 路强[1] 姜傥[1] 

机构地区：[1]中山大学附属第一医院检验医学科,广州510080

出　　处：《中华生物医学工程杂志》2015年第2期134-137,共4页Chinese Journal of Biomedical Engineering

摘　　要：目的：在PRShGRα质粒的基础上构建人糖皮质激素受体的M565R、C638S、L753F、A573Q、GR2439insA诱变表达受体，为研究糖皮质激素抵抗的分子机制打下基础。方法采用Stratagene公司生产的QuickChange定点诱变试剂盒对PRShGRα质粒进行诱变，并对诱变体的编码区全长进行测序。结果测序结果表明M565R、C638S、L753F、A573Q、GR2439insA定点诱变体均诱变成功，且没有碱基的错配、插入、缺失情况。结论成功构建M565R、C638S、L753F、A573Q、GR2439insA定点诱变体，为进一步研究受体功能变化奠定实验基础。Objective To construct mutants of human glucocorticoid receptors M565R,C638S, L753F,A573Q,GR2439insA based on PRShGRαplasmid,and to lay a foundation for further research on molecular mechanism of glucocorticoid resistance. Methods The Quickchange Site-Directed Mutagenesis Kit produced by Stratagene Company was used to induce mutagenesis on PRShGRαplasmid. Then,the full-length coding region of the mutants was sequenced. Results The DNA sequencing results showed that the site-directed mutants M565R,C638S,L753F,A573Q and GR2439insA were successfully constructed, without mismatch,insertion and deletion of the bases. Conclusion The successfully constructed site-directed mutants M565R,C638S,L753F,A573Q and GR2439insA can lay a foundation for further experiment on the changes of the receptor function.

关 键 词：受体糖皮质激素 质粒 诱变 定点 载体 

分 类 号：R644[医药卫生—外科学]