Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Investigation of Conserved Regions in Lipase Genes

Lipase 유전자의 보존적 영역 탐색

  • 이동근 (신라대학교 공과대학 생명공학과) ;
  • 김철민 (부산대학교 의과대학 의학과) ;
  • 김상진 (한국해양연구원 미생물연구실') ;
  • 이상현 (신라대학교 공과대학 생명공학과) ;
  • 이재화 (신라대학교 공과대학 생명공학과)
  • Published : 2003.10.01
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Abstract

For the investigation of conserved regions in lipase genes, 132 and 24 sequences were obtained from LED (Lipase Engineering Database) and COG (Clusters of Orthologous Groups of proteins), respectively. There was high diversity in lipase genes and peculiar amino acid sequences were conserved for each homologous family of LED. Similar conserved amino acid sequences were detected from COG0657 and Moraxella lipase 1 homologous group of LED. Although many studies have attempted to detect new lipase genes in procaryotes, they have been limited culturable bacteria. The importance of metagenome, including DNA from non-culturable bacteria, is known. Due to the high diversity, we assumed it might be possible to detect new lipase gene from metagenome. Due to the high diversity of nucleotide sequences in lipase genes, 10 primer sets were designed. Designed primer sets were inspected in BLAST of NCBI and they could amplify a part of the lipase gene from 222 to 713 bp. They can amplify 16.7%∼60.0% of each lipase homologous group which was 3.6 fold higher than each sets. They might offer a high probability of detecting new lipase genes, owing to high efficiency and the diversity of lipase genes.

Lipase 유전자들의 보존적 영역을 탐색하기 위해 LED(Lipase Engineering Database)와 COG (Clusters of Orthologous Groups of proteins)를 통하여 각각 132개와 24개의 서열을 얻어 분석하였다. Lipase 유전자의 염기서열은 아주 다양하였고 LED의 각 상동성그룹 (homologous family) 별로 독특한 아미노산 서열이 보존적인 것을 확인 할 수 있었다. COG0657에 속하는 lipase들은 LED의 Moraxella lipase 1 homologous group과 유사한 아미노산 보존적 영역이 있음을 확인하였다. 다양한 반응 조건에 적응하는 혹은 높은 활성을 갖는 새로운 lipase 유전자를 원핵생물에서 탐색하기 위하여 다양한 시도들이 수행되어 왔지만 그들은 모두 배양가능한 미생물에 국한되어 있었다. 배양할 수 없는 세균의 유전자원을 포함하는 메타게놈에 대한 유용성 역시 최근에 널리 인식되고 있다. Lipase 유전자의 다양성으로 보았을 때 메타게놈을 이용한 새로운 lipase 유전자를 찾는 작업도 가능할 것으로 사료되어 lipase유전자 일부를 (222∼713 bp) 증폭시키는 총 10개의 PCR primer를 설계하였으며 그 가능성을 NCBI의 BLAST를 통하여 검증하였다. Lipase 유전자의 염기서열은 아미노산 서열보다 아주 다양하여 비교적 많은 수의 primer set이 필요하였다. 각 primer set의 증폭효율은 각 LED group의 16.7%와 60.0% 사이였고 개별적인 primer set을 이용할 때 보다 3.6배 효율이 높은 것으로 드러났다. Lipase 유전자의 다양성은 설계된 primer set들을 이용하여 새로운 lipase 유전자를 검출할 가능성을 높이는 것으로 사료되었다.

Keywords

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