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J. Biochem, 1984, Vol. 96, No. 4 1205-1215
© 1984 Japanese Biochemical Society

research-article

Effect of the Replacement of pRoR Promoter of {lambda}dv Plasmid by lac Promoter on the Synthesis of DNA1

Tetsu HASE, Akihiko KITAMOTO and Yukito MASAMUNE

Faculty of Pharmaceutical Sciences, Kanazawa University Takaramachi, Kanazawa, Ishikawa 920

Derivatives of {lambda}dv whose pRoR promoter was replaced by lactose operon (lac promoter) were constructed and cloned in pBR322 plasmid. They were named pLOP-1, 2, and 3, and their structures are shown in Fig. 1. These plasmids were introduced into Escherichia coli (E. coli) lac iq and the effect of the inducer of lac promoter on the synthesis of plasmid DNA was examined. The synthesis of pLOP-2 or 3 DNA was strongly stimulated. pLOP-1, however, responded poorly to the inducer. Plasmids pLOP-2 and 3 were not segregated evenly into daughter cells after the induction and most of the progeny cells did not receive the plasmid. The synthesis of plasmid DNA after induction depended on the function of O and P genes and was inhibited by the addition of rifampicin and chloramphenicol. Most of the plasmid DNA synthesized after the induction cosedimented with the folded host chromosomal complex, suggesting an unusual structure of the DNA. When the inducer was removed, normal segregation of the plasmid resumed and the copy number of plasmid DNA decreased to the original level.

1Part of this work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.


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