한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제35권3호
  • /
  • Pages.177-183
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  • 2007
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  • 1598-642X(pISSN)
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  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Methylovorus sp. Strain SS1 DSM11726으로부터 rpoH 유전자의 클로닝과 염기서열 분석

Cloning and Nucleotide Sequence Analysis of the rpoH Gene from Methylovorus sp. Strain SS1 DSM11726

  • 엄치용 (한국기초과학지원연구원 서울센터 메타볼롬분석연구팀) ;
  • 송승은 (한국기초과학지원연구원 서울센터 메타볼롬분석연구팀) ;
  • 박미화 (한국기초과학지원연구원 서울센터 메타볼롬분석연구팀) ;
  • 김영민 (연세대학교 이과대학 생물학과)
  • Eom, Chi-Yong (Metabolome Analysis Team, Korea Basic Science Institute (KBSI)) ;
  • Song, Seung-Eun (Metabolome Analysis Team, Korea Basic Science Institute (KBSI)) ;
  • Park, Mi-Hwa (Metabolome Analysis Team, Korea Basic Science Institute (KBSI)) ;
  • Kim, Young-Min (Department of Biology, College of Science, Yonsei University)
  • 발행 : 2007.09.28
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초록

열충격 시그마인자를 코딩하는 유전자 rpoH가 결여된 돌연변이체 대장균(Escherichia coli satrain A7448)을, 메탄올 자화세균인 Methylovorus sp. strain SS1 DSM11726의 phagemid library로 형질전환 시켜서 $30^{\circ}C$에서 성장하는 Escherichia coli strain A7448 로부터 Methylovorus sp. strain SS1 DSM11726의 rpoH 유전자를 클로닝하고 그 염기서열을 분석하였다. 1,793-bp 염기서열 분석 결과 Methylovorus sp. strain SS1 DSM11726의 RpoH는 284개의 아미노산으로 이루어져 있었으며 예상된 분자량은 32,006, p1값은 5.79로 나타났으며, 동일계열의 ${\beta}$-proteobacteria에 속하는 세균들의 RpoH와 높은 상동성을 보여주었다. Methylovorus sp. strain SS1 DSM11726의 RpoH는 대장균의 RpoH의 기능을 대신할 수 있음을 보여주었다. 열충격 후 RpoH양은 15분까지 지속적으로 증가하다 20분 뒤 양이 감소하는 양상을 나타내었다. 이는 Methylovorus sp. strain SS1 DSM11726의 RpoH 단백질 역시 열에 의해 유도됨을 말해 준다.

Using complementation of RpoH deficient E. coli strain A7448, the rpoH gene encoding heat shock sigma factor 32 (${\sigma}^{32}$) from Methylovorus sp. strain SS1 DSM11726 was cloned and sequenced. Sequence analysis of a stretch of 1,796-bp revealed existence of an open reading frame encoding a polypeptide of 284 amino acid (32,006 dalton). Deduced amino acid sequence of the Methylovorus sp. strain SS1 RpoH showed that 59.6%, 39.1% and 51.4% identities with those of Nitrosomonas europaea (${\beta}$-proteobacteria), Agrobacterium tumefaciens ($\alpha$-proteobacteria) and E. coli (${\gamma}$-proteobacteria). The expression level of the functional ortholog of RpoH of Methylovorus sp. strain SS1 was increased transiently after heat induction, further indicating that it functions as a heat shock sigma factor.

키워드

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