넙치 (Paralichthys olivaceus) vasa 유전자의 full-length cDNA 분리 및 생식소 특이적 발현

다국어 초록 (Multilingual Abstract)

Until recent, primordial germ cells(PGCs) are recognized only by morphological observation, such as their large size and low nucleocytoplasmic ratio. For the molecular analysis of the reproduction, it is important to identify a specific marker of germ cell development and differentiation. The VASA, which was first identified in Drosophila, is reported as a germ-line cell specific marker gene in animals. Many other researches verified its germ cell specific expression during embryogenesis and gametogenesis. VASA is a member of the DEAD(Asp-Glu-Ala-Asp) protein family of ATP-dependent RNA helicase, and plays a critical role in germ-line cell linage. Vasa is expected to be an useful molecular marker for identification of PGCs in reproduction researches of aquaculture species. In this study, we isolated the vasa cDNA, and surveyed its gonad-specific expression in Paralichthys olivaceus. The full-length cDNA of P. olivaceus vasa cDNA was isolated and deduced amino acid sequence was compared to those of the other teleosts. It was 2,461bp long, consisted in 646 amino acids in its ORF region, 175bp of 5’-UTR, and 345bp 3’-UTR. Flounder VASA contained conserved DEAD box, arginine-glycine rich region, and other domains were found. Flounder vasa expressed strongly in the testis and ovary, confirming its property as an gonad-specific marker. These result is expected to be an useful marker for flounder reproduction research and related aquaculture industry.

목차 (Table of Contents)

• Abstract
• 서론
• 재료 및 방법
• 1. primers
• 2. Total RNA 분리
• 3. cDNA 합성
• 4. Polymerase Chain Reaction (PCR)
• 5. 형질전환
• 6. Sequencing 및 in-silico analysis
• 7. Rapid Amplification of cDNA Ends (RACE)
• 결과 및 고찰
• 1. 넙치 vasa full-length cDNA의 분리
• 2. 넙치 vasa 유전자의 생식소 특이적 발현
• 요약
• 사사
• 참고문헌