Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Effect of Ferrous Ion on the Formation of Exotoxin A from Pseudomonas sp. PY002 and Cloning of it's Gene

Pseudomonas sp. PY002에서 Exotoxin A의 생성에 미치는 철 이온의 영향과 Exotoxin A 유전자의 클로닝

  • Choi, Sun-Ah (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Kim, Ho-Sang (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Choi, Ji-Young (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Kang, Jeong-Suk (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Kim, Chun-Sung (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Kim, Duck-Lae (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Kim, Young-Ju (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Yeo, Myeong-Gu (Department of Biological Science, College of Natural Sciences, Chosun University) ;
  • Park, Yeol (Department of Biological Science, College of Natural Sciences, Chosun University)
  • 최선아 (조선대 자연과학대학 생물과학부) ;
  • 김호상 (조선대 자연과학대학 생물과학부) ;
  • 최지영 (조선대 자연과학대학 생물과학부) ;
  • 강정숙 (조선대 자연과학대학 생물과학부) ;
  • 김춘성 (조선대 자연과학대학 생물과학부) ;
  • 김덕례 (조선대 자연과학대학 생물과학부) ;
  • 김영주 (조선대 자연과학대학 생물과학부) ;
  • 여명구 (조선대 자연과학대학 생물과학부) ;
  • 박열 (조선대 자연과학대학 생물과학부)
  • Published : 1999.03.01
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Abstract

By SDS-polyacrylamide gel elcctrophoresis (SDS-PAGE) and immunoblot analysis that a protein with 66,000 daltons in size was recognized by P. aeruginosa anti-exotoxin A from P. sp. PY002. The yields of exotoxin A in P. sp. PY002 culture were influenced by the concentration of iron in the culture media. Increasing of the exotoxin A production and siderophore production was made slight increasing in the MKB medium. On the other hand, to clone the gene encoding the exoloxin A genomic library of P. sp. PY002 was constructed in pBluescript SK(+). From this library a exotoxin A homologous gene was isolated by immunological hybridization method using P. aemginosa anti-exoloxin A as a probe. Two putative clones were isolated and designated pETA23 and pETA42. Thc restriction analysis ol pETA42 demonstrated that thc 1760 bp insert contained one NcoI, PvuII, SstI, Kpnl and EcoRI site and three SmaI and HaeD sites.

Pseudomons sp. PY002의 exotoxin A 의 발현 양상을 관찰하기 위하여 P.aeruginosa PAO1의 anti-exotoxin A와 immunoblot hybridization을 실시한 결과 배지내에 유용 가능한 철이 고갈됨에 따라 exotoxin A의 발현양은 점차적으로 증가하는 양상을 보였으며, CAS 배지에 점적한 배양 상층액에서 siderophore의 발현양도 증가함을 보였다. P.sp.PY002 의 genomix library를 제조하여, exotoxin A를 분비하는 2개의 클론을 선별하려 pETA23과 pETA42 로 명명한 후, 반응성이 강한 Peta42를 선발하였다. pETA 42는 약 1.7kb 크기의 insert를 가지며, 양쪽 말단에 cloning site 인 pstI site 가 존재하며 2개의 NcoI, 1개의 PvuII, 1개의 SstI , 3개의 SmaI, 1개의 KpnI, 3개의 HaeII, 1개의 EcoRI site가 존재하였다.

Keywords

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