title: A novel inducible expression system for the functional study of toxic gene in bacteria.
abstract: The cloning and expression of toxic proteins in bacteria have posed a great challenge because of the leaky expression in inducible expression systems. Using artificial gene synthesis and clone screening methods, we identified a mutant T5 promoter, which significantly reduced leaky expression of lac operator. The mutant T5 promoter contains two T deletions at -35 region and may reduce promoter activity. A bacterial lethal gene, Φ174 lytic gene E, was successfully cloned in this system and expressed in the presence of isopropyl β-D-1-thiogalactopyranoside. The system is compatible with existing T5 inducible expression systems and can be used for the controlled expression of toxic proteins.
authors: Hubei-MOST KLOS and KLOBME, School and Hospital of Stomatology of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, People's Republic of China, gjhpp@hotmail.com.
authors: Jihua
authors: J
authors: Guo
authors: Rong
authors: R
authors: Jia
mesh_terms: D001430:Bacteriocins / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D007763:Lac Operon / Q000235:genetics; D009154:Mutation; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013862:Thiogalactosides / Q000378:metabolism
chemical_list: D001430:Bacteriocins; D013862:Thiogalactosides