Identification of the Gene Products Responsible for F Plasmid Partitioning

  • Kim, Sung-Uk (Bioproducts R.G., Genetic Engineering Research Institute, KIST) ;
  • Kazuo Nagai (Dept. of Bioengineering, Tokyo Institute of Technology) ;
  • Gakuzo Tamura (Dept. of Agricultural Chemistry, The Univ. of Tokyo,) ;
  • Yu, Ju-Hyun (Dept. of Food Engineering, Yonsei Univ.) ;
  • Bok, Song-Hae (Bioproducts R.G., Genetic Engineering Research Institute, KIST)
  • Published : 1993.12.01

Abstract

DNA subfragments, sopA, sopB and sopC which help to maintain the stability of an ori C plasmid, were derived from a mini-F plasmid DNA (EcoRI restriction fragment f5) after digestion with restriction endonuclease, and cloned in the vector plasmid pBR322. The recombinant plasmids obtained were introduced into E. coli KY7231 and E. coli CSR603 strains, and proteins specified by the mini-F fragments were analysed by SDS-PAGE. Two proteins encoded by the F fragments were detected, and their molecular weights were 41,000 and 37,000 daltons. Fluorography after one and two dimensional gel electrophoresis of the lysates showed that these two proteins had been overproduced in the cells which were allowed to incorporate radioactive amino acid after plasmid amplification by chloramphenicol treatment. The isoelectric points of sopA and sopB proteins were 6.6 and 7.0, respectively.

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