Johnston S, Lee J H, Ray D S
Gene. 1985;34(2-3):137-45. doi: 10.1016/0378-1119(85)90121-0.
Bacteriophage M13 gene II has been cloned in the plasmid expression vector pING1 and thereby placed under the control of the inducible araB promoter of Salmonella typhimurium. Upon induction with arabinose, gene II is transcribed as part of a polycistronic messenger RNA which initiates at the araB promoter. Subsequent translation of this message results in the coordinate, high-level expression of several proteins, including the gene II protein. Using this expression system, we have been able to overproduce gene II protein to a level of almost 15% of the total protein in Escherichia coli cells, thus providing an abundant source for its purification.
噬菌体M13基因II已被克隆到质粒表达载体pING1中,从而置于鼠伤寒沙门氏菌可诱导的araB启动子的控制之下。用阿拉伯糖诱导后,基因II作为多顺反子信使RNA的一部分被转录,该信使RNA从araB启动子起始。随后对该信使RNA的翻译导致几种蛋白质的协同、高水平表达,包括基因II蛋白。利用这个表达系统,我们已经能够在大肠杆菌细胞中将基因II蛋白过量生产到几乎占总蛋白15%的水平,从而为其纯化提供了丰富的来源。