桑树泛素延伸蛋白基因的原核表达

Prokaryotic Expression of Ubiquitin Extension Protein Gene in Mulberry

对前期克隆的桑树泛素延伸蛋白基因进行了生物信息学及原核表达分析,以pGEX-4T-1为载体,构建了桑树泛素延伸蛋白基因重组表达质粒,转入受体菌E.coli BL21。经IPTG诱导后,基因在大肠杆菌中得到有效表达。为提高表达效果,从不同诱导时间对目的蛋白表达条件进行优化,用SDS-PAGE电泳检测,结果表明在经37℃条件下,经1.0 mmol/L的IPTG诱导6 h后,原核表达效果明显提高。研究表明,桑树泛素延伸蛋白基因与其他植物具有较高的同源性,因此利用模式植物研究其功能及分子进化分析更加高效,为进一步探讨桑树的逆境生理机制提供了理论依据。

This paper analyzes bioinformatics and prokaryotic expression of ubiquitin extension protein gene in mulberry cloned at earlier stage,constructs recombinant expression plasmid of ubiquitin extension protein gene in mulberry with pGEX-4T-1 as the vector and switches to recipient bacterium E.coli BL21.After IPTG induction,the gene has effective expression in escherichia coli.To improve the expression effect,target protein expression conditions are optimized at different induction time and detected by SDS-PAGE electrophoresis.The result shows that prokaryotic expression effect improves significantly after 1.0 mmol/L IPTG induction for 6 h at 37 ℃.The research shows that ubiquitin extension protein gene in mulberry has high homology with other plants.Therefore,using model plant to study its functions and analyze molecular evolution is more efficient and provides theoretical basis for further discussion on physiological mechanism of mulberry in adverse situation.