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간흡충 cDNA library를 이용한 항원유전자의 검색 및 재조합항원의 생성

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 Isolation of a clonorchis sinensis antigen gene using clonorchis sinensis cDNA library and production of a corresponding recombina 
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[한글] 우리 나라에서 중요한 인체감염 기생충인 간흡충의 cDNA library를 제작하고 간흡충 감염 토끼혈청으로 선별하여 항원을 발현하는 유전자를 찾아 그 유전자의 특성 및 기능을 알아보고자 하였다. 감염 토끼혈청에 강하게 반응하는 clone중 약 38 kDa의 항원단백을 encoding하는 cDNA clone (pBCs44)을 선별하였으며 이 유전자(GenBank Accession number, AF136608)는 822 base pairs의 open reading frame을 가지고, 30개의 일정한 염기서열을 23번 반복하는 양상을 보였다. 이 유전자는 간흡충에서 2 kb의 mRNA로 발현되고 간흡충 genome에서 한 개의 copy로 존재함을 확인하였다, GRCSP로 명명된 이 항원단백은 충체의 난황선에서 발현됨을 면역형광항체법으로 확인하였다.
[영문] In order to isolate antigen genes of Clonorchis sinensis and characterize the function of antigenic molecule, we constructed C.sinensis cDNA expression library and screened the library with experimentally infected rabbit sera. Among these, one gene product which reacted strongly with rabbit sera was further investigated. Here we present the complete aminoacid sequences of this antigenic protein, as deduced from the nucleotide sequences of their relative cDNA (pBCs44, GenBank Accession number AF136608). The nucleotide sequence of the cDNA revealed the presence of an open reading frame of 822 base pairs which had 23 tandemly arranged 30 nucleotide-long repeats encoding bacterial fusion protein of 38 kDa. The insert cDNA from the pBCs44 clone hybridized to a RNA of about 2 kb in a northern blot analysis. Southern blot analysis indicated that pBCs44 gene was likely to exist as a single copy in the genome of C. sinensis. Immunofluorescence studies using specific anti-fusion protein sera revealed that the antigen was specifically expressed in the vitellaria.
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