Enhanced Activity of Cytidine Deaminase by Gene Family Shuffling.

Gene Family Shuffling을 이용한 Cytidine Deaminase 활성 증가

  • Hong, Sik (Department of Microbiology, Kyungpook National University) ;
  • Kim, Kyung-Dong (Department of Microbiology, Kyungpook National University) ;
  • Song, Bang-Ho (Department of Biology Education, Kyungpook National University) ;
  • Jung, Kyung-Hwa (Genomics Laboratory, Amicogen Inc.) ;
  • Kim, Sa-Yeol (Department of Microbiology, Kyungpook National University)
  • 홍식 (경북대학교 미생물학과) ;
  • 김경동 (경북대학교 미생물학과) ;
  • 송방호 (경북대학교 생물교육과) ;
  • 정경화 (아미코젠(주) 제노믹스연구실) ;
  • 김사열 (경북대학교 미생물학과)
  • Published : 2002.12.01


A family shuffling associating PCR-based and in vitro recombination and expression in Escherichia coli cdd mutant was carried out. Two cdd genes encoding cytidine deaminases (CDase) from thermophilic Bacillus caldolyticus and B. stearothermophilus were shuffled. Around 150 viable mutant colonies screened on AB minimal medium without uracil by E. coli cdd complementation were selected for cytidine deaminase assay and 4 candidates (SH1067, SH1077, SH1086, and SH1118) were chosen for the detailed study. The nucleotide sequence analyses of 4 selected mutants revealed that they have several point mutations and recombinations. Surprisingly, the SH 1067 showed 770 fold more specific CDase activity at $80^{\circ}C$ than that of T101 from parental B. stearothermophilus.


Bacillus caldolyticus;B. stearothermophilus;cytidine deaminase;family shuffling.


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