Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Nucleotide Sequence Analysis of the rpoH Gene from Methylovorus sp. Strain SS1 DSM11726

Methylovorus sp. Strain SS1 DSM11726으로부터 rpoH 유전자의 클로닝과 염기서열 분석

  • 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)
  • 엄치용 (한국기초과학지원연구원 서울센터 메타볼롬분석연구팀) ;
  • 송승은 (한국기초과학지원연구원 서울센터 메타볼롬분석연구팀) ;
  • 박미화 (한국기초과학지원연구원 서울센터 메타볼롬분석연구팀) ;
  • 김영민 (연세대학교 이과대학 생물학과)
  • Published : 2007.09.28
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Abstract

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.

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.

Keywords

References

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