安徽工程大学学报2016，Vol.31Issue(2)：11-15,5.
pGEX－4T－2－A表达系统的构建及其融合蛋白GST－PlaA的表达优化
Construction of expressive system pGEXG4TG2GA and the expression and optimization of its GSTGPlaA fusion protein
摘要
Abstract
To construct a recombinant plasmid pGEX-4T-2-A for GST-PlaA fusion protein expression. PCR was used to amplify phopholipase A1 gene from Serratia marcescens PL-06 named plaA and insert-ed into the pGEX-4T-2 vector.The recombinant plasmid was transformed to Escherichia coli JM109 strains to screen the positive recombinant clones and then sent to be sequenced.The correct sequenced re-combinant plasmid was transformed to a Escherichia coli BL2 1 (DE3 )PLysS strains and the GST-PlaA fusion protein expression after IPTG induction was confirmed by SDS-PAGE analysis.Its cultivation con-ditions was also optimized.The plasmid pGEX-4T-2-A was constructed successfully,the GST-PlaA fu-sion protein expressed in the form of inclusion bodies.While the OD was 0.8 and induced at 37 ℃ for 8 h with 0.5 mM IPTG,the GST-PlaA fusion protein expression was at the highest level.This experiment es-tablished the foundation for the purification and function of phospholipase A1 protein.关键词
磷脂酶 A1/融合蛋白/SDS-PAGE/表达条件Key words
phospholipase A1/fusion protein/SDS-PAGE/expression conditions分类
生物科学引用本文复制引用
刘开放,薛正莲,王洲..pGEX－4T－2－A表达系统的构建及其融合蛋白GST－PlaA的表达优化[J].安徽工程大学学报,2016,31(2):11-15,5.基金项目
国家自然科学基金资助项目(31471615,31501465) （31471615,31501465）
