A new idea for simple and rapid monitoring of gene expression: requirement of nucleotide sequences encoding an TV-terminal HA tag in the T7 promoter-driven expression in E. coli

摘要

Mammalian expression vectors are used to overexpress genes of interest in mammalian cells. High temperature requirement protein Al (HtrAl), used as a specific target, was expressed from the pHA-M-HtrAl plasmid in HEK293T cells, inducing cell death. Expression of HtrAl was driven by the pHA-M-HtrAl mammalian expression vector in E. coli resulting in growth suppression of E. coli in an HtrAl serine protease-dependent manner. By using various combinations of promoters, target genes and A/-terminal tags,the T7 promoter and N-terminal HA tag in the mammalian expression vector were shown to be responsible for expression of target genes in E. coli. Thus the pHA-M-HtrAl plasmid can be used as a novel, rapid pre-test system for expression and cytotoxicity of the specific target gene in E. coli before assessing its functions in mammalian cells.