Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Cloning and Regulation of Schizosaccharomyces pombe Gene Encoding Ribosomal Protein S20

  • Lee, Yoon-Jong (Division of Life Sciences, Kangwon National University) ;
  • Kim, Kyunghoon (Division of Life Sciences, Kangwon National University) ;
  • Park, Eun-Hee (College of Pharmacy, Sookmyung Women′s University) ;
  • Ahn, Ki-Sup (Department of Environmental Management, Chonan College of Foreign Studies) ;
  • Kim, Daemyung (Department of Genetic Engineering, Congju University) ;
  • Lim, Chang-Jin (Division of Life Sciences, Kangwon National University)
  • Published : 2001.03.01
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Abstract

A cDNA clone encoding the ribosomal protein S20 has been isolated from the Schizosaccharomyces pombe cDNA library by colony hybridization. The insert contained in the original plasmid pYJ10 was transferred intro shuttle vector pRS316 generate plasmid pYJll. The dDNA insert of plasmid pYJll, contains 484 nucleotides and encodes a protein of 118 amino acids with a calculated mass of 13,544 daltons. The deduced amino acid sequence of S. pombe ribosomal protein S20 is very homologous with fruit fly, rat, and budding yeast counterparts. It is also homologous with Xenopus S22 ribosomal protein. S. pombe ribosomal protein S20 appears to be relatively hydruphobic except the C-terminal region. The 728 bp upstream region of the S20 gene was amplified from chromosomal DNA and transferred into the BamHI/EcoRI site of the promoterles $\beta$-galactosidase gene of the vector YEp357R, which resulted in fusion plasmid pYS20. The synthesis of $\beta$-galactosidase from the fusion plasmid appeared to be the highest in the mid-exponential phase. The S. pombe cells with the fusion plasmid grown at 35$\^{C}$ gave lower $\beta$-galactosidase activity than the cells grown at 30$\^{C}$. Computer analysis showed the consensus sequence CAGTCACA in the upstream regions of various ribosomal protein genes in S. pombe, which would be involved in the coordinated expression of small ribosomal proteins.

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