质粒
大肠杆菌
紫胶操纵子
重组DNA
包涵体
克隆(编程)
化学
靶蛋白
表达式向量
分子生物学
变性(裂变材料)
生物化学
生物
计算生物学
基因
核化学
程序设计语言
计算机科学
作者
Herui Gao,Xianghui Qi,Darren J. Hart,Song Gao,Hongling Wang,Shuangyan Xu,Yifeng Zhang,Xia Liu,Yifei Liu,Yi An
标识
DOI:10.1021/acs.jafc.8b01960
摘要
We have constructed novel plasmids pANY2, pANY3, and pANY6 for flexible cloning with low false positives, efficient expression, and convenient purification of proteins. The pANY2 plasmid can be used for efficient isopropyl-β-d-thiogalactoside (IPTG) induced protein expression, while the pANY3 plasmid can be used for temperature-induced expression. The pANY6 plasmid contains a self-cleaving elastin-like protein (ELP) tag for purification of recombinant protein by simple ELP-mediated precipitation steps and removal of the ELP tag by self-cleavage. A urea-based denaturation and refolding processes for renaturation of insoluble inclusion bodies can be conveniently integrated into the ELP-mediated precipitation protocol, removing time-consuming dialysis steps. These novel vectors, together with the described strategies of gene cloning, protein expression, and purification, may have wide applications in biosciences, agricultural, food technologies, and so forth.
科研通智能强力驱动
Strongly Powered by AbleSci AI