견사곤충에서 Mariner Transposase-like Element의 분자적 동정

Molecular identification of Mariner Transposase-like Element from Four silkmoths

  • 이진성 (생명공학연구소 유전체사업단) ;
  • 황재삼 (농촌진흥청 잠사곤충연구소) ;
  • 김용성 (생명공학연구소 유전체사업단) ;
  • 서동상 (성균관대학교 유전공학과)
  • 발행 : 1998.08.01

초록

누에, 멧누에, 천잠 및 작잠에서의 형질전환용 vector 개발을 위한 첫단계로 이미 인간을 포함한 여러 곤충에서 그 존재가 확인된 mariner transposable element가 이들 견사 곤충에 존재하는지의 여부를 탐색하기 위해서 이미 보고된Drosophila mauritiana와 Hyalophora ceropia의 mariner transposase 유전자의 고도로 보존성이 높은 부위에 대한 degenerctive primer pair를 제작하여 이들의 존재 여부를 탐색하였다. 결과적으로 예상되는 약 500bp의 PCR product가 관찰됨에 따라서 모든 견사곤충의 genome에는 maminer-like element(MLE)가 존재한다는 것을 간접적으로 확인할 수 있었다. 이미 cloning된 pBmoMAR를 probe DNA로하여 southern hybridization을 수행한 결과에서도 확인 할 수 있었으며, 각 견사층의 genome내에 high copy로 존재한다는 것을 추정할 수 있었다. 따라서 이와 같은 결과는 견사곤충에서 MLE가 vector system으로써 개발될 수 있는 기초자료로 제공될 수 있을 것이다.

As a first step for developing universal genetic transformation vector of dilkmoths., we identified the presence of mariner-like element(MLE) which is one of transposable element discovered from many insects to human species, from Bombyx mori, Bombyx mandarina, Antherae yammamai and Antherae pernyi. We used a degenerative primer pair designed from a transposase gene of Drosophila mauritiana and Hyalophora ceropia MLE. As results, major PCR product of 500bp expected as a part of transposase of MLE was detected from all the slkmoths used of this study using these primer. And hybridization assay using pBmoMAR as a probe DNA that was previously cloned from Bombyx mori by the same primer pair, confirmed the presence of MLE from all the silkmoths. This assay showed also that the endogenous MLE in genome of the silkworm is present as high copy number unlikely Drosophila mauritiana which has 10-20 copy number. This data will be a fundamental genetic information for developing mariner-derived vector to transform the silkmoths and other useful insects.

키워드

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