Cloning of Autoregulator Receptor Gene form Saccharopolyspora erythraea IFO 13426

Saccharopolyspora erythraea IFO 13426으로부터 Autoregulator Receptor Protein Gene의 Cloning

  • 김현수 (계명대학교 자연과학대학 미생물학과) ;
  • 이경화 (계명대학교 자연과학대학 미생물학과) ;
  • 조재만 (계명대학교 자연과학대학 미생물학과)
  • Published : 2003.06.01

Abstract

For screening of autoregulator receptor gene from Saccharopolyspora erythraea, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHI site of pUC19 and transformed into the E. coli DH5$\alpha$. The isolated plasmid from transformant contained the fragment of 120 bp, which was detected on 2% gel after BamHI treatment. The insert, 120 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridization using Saccha. erythraea chromosomal DNA were performed with the insert as probe. The plasmid (pEsg) having 3.2 kbp SacI DNA fragment from Saccha. erythraea is obtained. The 3.2 kbp SacI DNA fragment was sequenced by the dye terminator sequencing. The nucleotide sequence data was analyzed with GENETYX-WIN (ver 3.2) computer program and DNA database. frame analyses of the nucleotide sequence revealed a gene encoding autoregulator receptor protein which is a region including KpnI and SalI sites on 3.2 kbp SacI DNA fragment. The autoregulator receptor protein consisting of 205 amino acid was named EsgR by author. In comparison with known autoregulator receptor proteins, homology of EsgR showed above 30%.

Keywords

Saccharopolyspora erythraea;\gamma-butyrolactone autoregulator;receptor protein;erythromycin

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