Molecular Cloning of Escherichia coli cdd Gene Encoding Cytidine/Deoxycytidine Deaminase.

Escherichia coli의 시티딘/디옥시시틴딘 디아미나제를 코드하는 cdd 유전자의 클로닝

  • 권택규 (경북대학교 자연과학대학 미생물학과) ;
  • 김태호 (경북대학교 자연과학대학 미생물학과) ;
  • 황선갑 (경북대학교 자연과학대학 미생물학과) ;
  • 김종국 (경북대학교 자연과학대학 미생물학과) ;
  • 송방호 (사범대학 생물교육과) ;
  • 홍순덕 (사범대학 생물교육과)
  • Published : 1990.12.01

Abstract

We have cloned the cdd gene from E. coli C600 using (cdd-) as a host. From the sequenced promoter region of E=, coli cdd gene which has been determined by Valentin-Hansen P. (1985), we synthesized the 23 mer oligonucleotides corresponding to the transcription initiation region and used as a probe for cloning of the cdd gene by Southern blotting. The isolated fragments in the blotting were introduced to the colony hybridization after transforming it into the E. coli JF611 (cdd-, pyr double mutant), and we identified the hybridized band at 27 kb long. From the original insert of 27 kb fragment in theBamHI site of pBR322, the 5.3 kb fragment containing the cdd gene was isolated by subsequent deletion and subeloning. From the derived plasmid pTK509, further deletion and subcloning were performed and clarified that the cdd gene was located in the 2.1 kb of SaZI/DraI segment in the insert of pTK605. The polypeptide encoded by the cloned DNA was appeared to be a molecular mass of 33,000.

Keywords

cdd gene;cytidine/deoxycytidine deaminase.