Noguchi T, Takahashi H
Research Laboratories, Yamasa Shoyu Co., Ltd., Chiba, Japan.
Agric Biol Chem. 1991 Oct;55(10):2497-506.
We established an efficient system for a high-level production of foreign gene products in Escherichia coli using a trans-activated gene expression under the control of a phage T4 regulatory signal cloned in a plasmid by T4 phage infection. The transcriptional and translational signal of the T4 uvsY gene cloned in a plasmid was fused translationally with the coding region of the lacZ gene. When E. coli cells carrying the uvsY-lacZ plasmid were infected with cytosine-substituting T4 phage at a multiplicity of infection of 5, the amount of beta-galactosidase increased about 2-fold (trans-activation) over that without phage infection. We examined conditions for the high-level production of a trans-activated gene product. We found that a large number of T4-infected cells in a lysis-inhibition state could be obtained by a low multiplicity of infection with cytosine-substituting T4 phage. Thus it is now possible to attain a high yield of the trans-activated gene products. We discuss the advantage of the trans-activated T4 uvsY regulatory signal for production of foreign products.
我们建立了一种高效系统，可利用噬菌体T4调控信号控制下的反式激活基因表达，在大肠杆菌中高水平生产外源基因产物。通过T4噬菌体感染，将克隆在质粒中的噬菌体T4调控信号用于控制反式激活基因表达。克隆在质粒中的T4 uvsY基因的转录和翻译信号与lacZ基因的编码区进行翻译融合。当携带uvsY-lacZ质粒的大肠杆菌细胞以感染复数为5被胞嘧啶取代的T4噬菌体感染时，β-半乳糖苷酶的量比未感染噬菌体时增加了约2倍（反式激活）。我们研究了反式激活基因产物高水平生产的条件。我们发现，通过用胞嘧啶取代的T4噬菌体进行低感染复数感染，可以获得大量处于裂解抑制状态的T4感染细胞。因此，现在有可能获得高产的反式激活基因产物。我们讨论了反式激活的T4 uvsY调控信号在生产外源产物方面的优势。
