Noguchi T, Takahashi H
Research Laboratories, Yamasa Shoyu Co., Ltd., Chiba, Japan.
Agric Biol Chem. 1991 Oct;55(10):2507-13.
A novel expression system was developed for the high level production of a labile protein in Escherichia coli. The regulatory signal of bacteriophage T4 uvsY gene was fused in frame with the coding region of human ventricular myosin alkali light chain (VLC1) gene. Expression from the regulatory signal was enhanced and continued in a lysis-inhibition state by infection with a cytosine-substituting T4 phage mutant. VLC1 protein was produced at a low level without infection because of its instability in the cells. Although the productivity was partly improved in a lon-deficient mutant without infection, it was improved about 100-fold with T4 phage infection. T4 phage produces protease inhibitor(s) (pin gene product) against proteases of host cell including the lon gene product (protease La).
开发了一种新型表达系统，用于在大肠杆菌中高水平生产一种不稳定蛋白。将噬菌体T4 uvsY基因的调控信号与人心室肌球蛋白碱性轻链（VLC1）基因的编码区读框融合。通过用胞嘧啶替代的T4噬菌体突变体感染，调控信号的表达得到增强，并在裂解抑制状态下持续。由于VLC1蛋白在细胞中不稳定，未感染时其产生水平较低。虽然在未感染的lon缺陷突变体中生产力有所部分提高，但在T4噬菌体感染时提高了约100倍。T4噬菌体产生针对宿主细胞蛋白酶（包括lon基因产物蛋白酶La）的蛋白酶抑制剂（pin基因产物）。
