小鼠Klf4基因的原核表达研究
The Research of Prokaryotic Expression of Krüppel-like Factor 4 in Mouse
孟书燕 1郭杰 1高庚渠 1刘改霞 1马威1
作者信息
- 1. 河南质量工程职业学院,河南平顶山 467000
- 折叠
摘要
[目的]对小鼠Klf4基因重组蛋白进行原核表达分析.[方法]利用双酶切从重组质粒pMXs-Klf4上回收小鼠Klf4基因片段,克隆入pET-41a(+)载体后转化BL21大肠杆菌,用IPTG诱导表达,探讨诱导表达最佳浓度和时间,最后采用SDS-PAGE对重组蛋白进行鉴定及分析.[结果]重组质粒Klf4-pET-41a(+)在IPTG诱导下可表达与预期相符的约为51 ku的KLF4蛋白;经IPTG刺激后重组蛋白表达增多,以0.8 mmol/L IPTG诱导6h为佳;重组蛋白以包涵体形式存在于大肠杆菌BL21中.[结论]小鼠Klf4基因可在原核细胞中表达.
Abstract
[Objective] Prokaryotic expression analysis of recombinant protein of Klf4 gene in mice was conducted.[Method] Klf4 gene segments were recovered from recombinant plasmid pMXs-Klf4 by double enzyme digestion,then they were cloned into pET-41a (+) vector and transformed into E.coli BL21.The expression was induced by IPTG,the optimal concentration and time was explored,finally,SDS-PAGE was adopted to identify and analyze recombinant plasmid.[Result] Induced by IPTG,recombinant plasmid Klf4-pET-41a(+) can be expressed in accordance with the expected protein about 51 ku;after IPTG stimulation,the expression of recombinant protein was increased,and 0.8 mmol/L IPTG induction for 6 h was optimal;the recombinant protein was expressed in the form of inclusion bodies in E.coli BL21.[Conclusion] The mouse Klf4 gene can express in prokaryotic cells.
关键词
小鼠/Klf4基因/原核表达/大肠杆菌Key words
Mouse/Klf4 gene/Prokaryotic expression/E.coli引用本文复制引用
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据