pKT230 벡터를 이용한 Pseudomonas sp. P20의 2,3-Dihydroxybiphenyl Dioxygenase 유전자의 클로닝

  • 김지영 (충북대학교 자연과학대학 미생물학과) ;
  • 김치경 (충북대학교 자연과학대학 미생물학과) ;
  • 가종억 (서울대학교 분자미생물학연구센터) ;
  • 민경희 (서울대학교 분자미생물학연구센터) ;
  • 박용근 (서울대학교 분자미생물학연구센터)
  • Published : 1996.12.01

Abstract

Pseudomonas sp. P20 isolated from the polluted environment is capable of degrading biphenyl and 4-chlorobiphenyl. The pcbABCD genes responsible for degradation of biphenyl and 4-chlorobiphenyl were cloned using pBluescript SK(+) from the chromosomal DNA of Pseudomonas sp. P20 to construct pCK1 and pCK102, harbouring pcbABCD and pcbCD, respectively. The 2, 3-DHBP dioxygenase gene, pcbC, was cloned again from pCK102 by using pKT230 which is known as a shuttle vector and pKK1 hybrid plasmid was constructed. The E. coli KK1 transformant obtained by transforming the pKK1 into E. coli XL1-Blue showed 2, 3-DHBP dioxygenase activity. The specific 2, 3-DHBP dioxygenase activity of E. coli KK1 was similar to that of the E. coli CK102, but much higher than those of the natural isolates, Pseudomonas sp. DJ-12 and Pseudomonas sp. P20.

Keywords

References

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