Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Cloning and Characterization of BTG-1 Gene from Pacific Oyster (Crassostrea gigas)

참굴(Crassostrea gigas)의 BTG1 유전자의 특성

  • Chung, In Young (Department of Microbiology, Pukyong National University) ;
  • Oh, Jeong Hwan (Institute of Fisheries Science, College of Fisheries Science, Pukyong National University) ;
  • Song, Young Hwan (Department of Microbiology, Pukyong National University)
  • 정인영 (국립부경대학교 미생물학과) ;
  • 오정환 (국립부경대학교 수산과학연구소) ;
  • 송영환 (국립부경대학교 미생물학과)
  • Received : 2017.02.08
  • Accepted : 2017.02.27
  • Published : 2017.04.30
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Abstract

BTG 1 (B-cell translocation gene 1) gene was first identified as a translocation gene in a case of B-cell chronic lympocytic leukemia. BTG1 is a member of the BTG/TOB family with sharing a conserved N-terminal region, which shows anti-proliferation properties and is able to stimulate cell differentiation. In this study, we identified and characterized the pacific oyster Crassostrea gigas BTG1 (cg-BTG1) gene from the gill cDNA library by an Expressed Sequence Tag (EST) analysis and its nucleotide sequence was determined. The cg-BTG1 gene encodes a predicted protein of 182 amino acids with 57% 56% identities to its zebrafish and human counterparts, and is an intron-less gene, which was confirmed by PCR analysis of genomic DNA. Maximal homologies were shown in conserved Box A and B. The deduced amino acid sequence shares high identity with other BTG1 genes of human, rat, mouse and zebrafish. The phylogenic analysis and sequence comparison of cg-BTG1 with other BTG1 were found to be closely related to the BTG1 gene structure. In addition, the predicted promoter region and the different transcription-factor binding site like an activator protein-1 (AP-1) response element involved in negative regulation and serum response element (SRE) were able to be identified by the genomic DNA walking experiment. The quantitative real-time PCR analysis showed that the mRNA of cg-BTG1 gene was expressed in gill, heart, digestive gland, intestine, stomach and mantle. The cg-BTG1 gene was expressed mainly in heart and mantle.

BTG1 (B-cell translocation gene 1)은 APRO family (anti-proliferative protein family)에 속하며, 이들은 공통의 생물학적 기능은 세포증식을 억제하는 것으로 알려져 있다. 본 연구에서, 굴의 gill cDNA library를 random sequencing을 통한 EST 분석과정에서 BTG1 clone을 확보하였으며, 분자생물학적 특성을 조사하였다. 굴의 BTG1은 182 개의 아미노산으로 구성되며, zebrafish와 57%, human과 56%의 상동성을 나타냈으며, 사람이나 설치류와 달리 ORF (open reading frame) 내에 intron이 존재하지 않았다. Genomic DNA walking을 통해 굴의 BTG1의 predicted promoter를 확인하였으며, 분석결과 AP-1 element와 SRE (serum response element) 부위가 존재하였으며, 5'flanking region에 cAMP response element (CRE) 부위가 확인되었다. 굴의 BTG1의 조직별 유전자발현 수준을 확인하기 위해 real-time PCR을 수행하였으며, 6 개 조직 모두에서 BTG1의 유전자발현이 나타났으며, 그 중에서 heart와 mantle에서 높은 수준의 mRNA 발현을 확인할 수 있었다.

Keywords

References

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