高赖氨酸蛋白基因在大肠埃希菌中的表达

从四棱豆中克隆高赖氨酸蛋白基因wblys,通过PCR扩增wblys片段,转入原核表达载体pGEX-4T-1,构建pGEX-4T-1/wblys大肠埃希菌工程菌,表达重组蛋白,IPTG诱导后,发现细菌全蛋白在44 ku(含GST标签)处多出1条明显条带。HPLC检测赖氨酸含量。诱导后菌体总赖氨酸含量比正常菌体提高15.84 mg/g。在大肠埃希菌中高效表达植物源高赖氨酸蛋白基因,为该基因在益生菌中表达提供研究工作基础。

Expression of a Lysine-Rich Gene in E. coli

A lysine-rich protein gene wblys was cloned from winged bean,a fragment of it was amplified through PCR and transferred into prokaryotic expression vector pGEX-4T-1,the recombinant plasmid pGEX-4T-1/wblys were constructed and then transferred to E.coli,the recombinant protein were expressed.Upon IPTG induction,the recombinant pGEX-4T-1/wblys produced a new protein with an apparent MW of 44 ku（containing GST marker） with an additionally obvious band.The content of lysine was detected by HPLC.After the induction the content of lysine of the bacteria increased by 15.84 mg/g as compared with the normal one.The high-effective expression of a plant-source lysine-rich gene in E.coli,providing a foundation for the study on its expression in probiotics.