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.
出处
《中华生物医学工程杂志》
CAS
2015年第2期134-137,共4页
Chinese Journal of Biomedical Engineering
关键词
受体糖皮质激素
质粒
诱变
定点
载体
Receptors,glucocorticoid
Plasmids
Mutagenesis,Site-directed
Vectors
