期刊论文

On Expression of Human KLK7 Gene and Its Effects on Cell Proliferation and Apoptosis

中文

衡阳师范学院学报

1673-0313

2022

43

3

89-94

10.3969/j.issn.1673-0313.2022.03.012

2019JJ60022:湖南省自然科学联合基金项目 2020JJ6042:湖南省自然科学联合基金项目

本校为第一机构

本研究旨在对人激肽释放酶7 （KLK7）基因进行原核与真核表达，研究其功能，并为后续KLK7新型生物抑制剂的研制提供基础材料。采用PCR扩增KLK7基因，将其克隆至原核表达载体pGEX4T-1和真核表达载体pEGFP-C1，构建重组表达质粒pGEX4T-1-KLK7和pEGFP-C1-KLK7，分别转化Transetta （DE3）和DH5α感受态细胞，筛选阳性菌株，用IPTG诱导重组KLK7蛋白（rKLK7）表达，SDS-PAGE和Western blot分析表达情况。将pEGFP-C1-KLK7转染人角质形成细胞HaCaT，观察KLK7在细胞中的定位，检测KLK7基因过表达对角质化细胞增殖的影响。KLK7基因开放阅读框为762 bp，编码254个aa。原核系统表达的rKLK7分子量为53.5 kDa...

The purpose of this study was to investigate the prokaryotic and eukaryotic expression of human kallikin-releasing enzyme 7(KLK7)gene,and to provide basic materials for the subsequent development of novel biological inhibitors of KLK7.KLK7 gene was amplified by PCR and cloned into prokaryotic expression vector pGEX4T-1 and eukaryotic expression vector PEGFP-C1.Recombinant expression plasmids pGEX4T-1-KLK7 and pEGFP-C1-KLK7 were constructed and transformed into Transetta(DE3)and DH5αcompetent cell,respectively.The recombinant KLK7 protein(rKLK7...