Utilization of trans-activated T4 uvsY regulatory signal for a high yield of cloned gene products.
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.
Noguchi
T
T
Research Laboratories, Yamasa Shoyu Co., Ltd., Chiba, Japan.
Takahashi
H
H
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003596:Cytosine / Q000502:physiology; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D013604:T-Phages / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics*
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D003596:Cytosine