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Position: Home > Articles > The Research of Prokaryotic Expression of Krüppel-like Factor 4 in Mouse Journal of Anhui Agricultural Sciences 2016 (12) 133-135

小鼠Klf4基因的原核表达研究

作  者:
孟书燕;郭杰;高庚渠;刘改霞;马威
单  位:
河南质量工程职业学院
关键词:
小鼠;Klf4基因;原核表达;大肠杆菌
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p MXs-Klf4上回收小鼠Klf4基因片段,克隆入p ET-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-p ET-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6 h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。
译  名:
The Research of Prokaryotic Expression of Krüppel-like Factor 4 in Mouse
作  者:
MENG Shu-yan;GUO Jie;GAO Geng-qu;MA Wei;Henan Quality Polytechnic;
关键词:
Mouse;;Klf4 gene;;Prokaryotic expression;;E;;coli
摘  要:
[Objective] Prokaryotic expression analysis of recombinant protein of Klf4 gene in mice was conducted.[Method] Klf4 gene segments were recovered from recombinant plasmid p MXs-Klf4 by double enzyme digestion,then they were cloned into p ET-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-p ET-41a(+) can be expressed in accordance with the expected protein about 51 ku;after IPTG stimulation,the expression of recombinant protein was increased,and0.8 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.

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