Bhandari P, Gowrishankar J
Centre for Cellular and Molecular Biology, Hyderabad, India.
J Bacteriol. 1997 Jul;179(13):4403-6. doi: 10.1128/jb.179.13.4403-4406.1997.
Salt-induced overexpression of genes cloned downstream of the phage T7 phi10 promoter was demonstrated in an Escherichia coli strain (GJ1158) which carries a single chromosomally integrated copy of the gene for phage T7 RNA polymerase under transcriptional control of the cis-regulatory elements of the osmoresponsive proU operon. Plasmids that have been constructed to obtain overproduction of individual target gene products in strain BL21(DE3) (by addition of isopropyl-beta-D-thiogalactopyranoside as an inducer) can directly be transformed into GJ1158. The NaCl induction regimen was also shown to be associated with a decreased propensity for sequestration of overexpressed target proteins within insoluble inclusion bodies.
在大肠杆菌菌株(GJ1158)中证实,盐诱导了在噬菌体T7 phi10启动子下游克隆的基因的过表达,该菌株携带一个整合在染色体上的噬菌体T7 RNA聚合酶基因拷贝,其受渗透反应性proU操纵子的顺式调控元件的转录控制。为在菌株BL21(DE3)中过量生产单个目标基因产物而构建的质粒(通过添加异丙基-β-D-硫代半乳糖苷作为诱导剂)可直接转化到GJ1158中。还表明,NaCl诱导方案与减少过表达的目标蛋白在不溶性包涵体内隔离的倾向有关。