Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
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Characterization of Mud Loach (Misgurnus mizolepis) Apolipoprotein A-I: cDNA Cloning, Molecular Phylogeny and Expression Analysis

미꾸라지(Misgurnus mizolepis) Apolipoprotein A-I cDNA의 구조, 분자계통 및 발현 특징 분석

  • Lee, Youn-Ho (National Fisheries Research and Development Institute) ;
  • Noh, Jae-Koo (National Fisheries Research and Development Institute) ;
  • Kim, Keun-Yong (Department of Aquaculture, Pukyong National University) ;
  • Cho, Young-Sun (Department of Aquaculture, Pukyong National University) ;
  • Nam, Yoon-Kwon (Department of Aquaculture, Pukyong National University) ;
  • Kim, Dong-Soo (Department of Aquaculture, Pukyong National University)
  • Published : 2007.02.25
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Abstract

Full length complementary DNA encoding apolipoprotein A-I (apoA-I) was isolated and characterized in mud loach (Misgurnus mizolepis). Mud loach apoA-I cDNA encoding 24 bp of 5'-untranslated region (UTR), 762 bp of single open reading frame (ORF) consists of 254 amino acids and 293 bp of 3'-UTR excluding stop codon and poly (A+) tail. Two overlapping polyadenylation signals (AATAAAATAAA) was found 9 bp prior to the poly (A+) tail. Mud loach apoA-I represented considerable homology to those from other teleost species at amino acid level with conserving common features of vertebrate apoA-I. Molecular phylogenetic analysis inferred the phylogenetic hypothesis that was generally in accordance with the previous taxonomic relationship. Apolipoprotein A-I mRNA was detected in various tissues, but the mRNA levels were quite varied depending on tissues based on semi-quantitative RT-PCR. Liver and brain showed the significantly higher levels of apoA-I transcripts than other tissues. mRNA expression of apoA-I was quite low in very early stage of embryonic development, however dramatically enhanced from 8 hours post fertilization. This increased mRNA level was retained consistently up to 14 days post hatching.

우리나라 주요 담수어종인 미꾸라지(Misgurnus mizolepis)로 부터 apolipoprotein A-I (apoA-I) cDNA를 분리하고 그 구조, 분자 계통 및 발현 특징을 분석하였다. 미꾸라지 apoA-I cDNA는 254개의 아미노산을 암호화하고 있는 762 bp의 ORF를 포함하고 있었으며 아울러 24 bp의 5'UTR 및 293 bp의 3'UTR(종결 코돈 및 poly A tail 제외)를 갖고 있었다. 미꾸라지 apoA-I은 여타 척추동물 apoA-I과의 다중배열 시 염기서열에서는 많은 차이를 나타내었지만 단백질의 구조적 특징은 높은 상동성을 보였고, 또한 척추동물의 apoA-I들과의 분자계통을 분석한 결과, 종래 알려진 분류학적 위치와 비교적 잘 일치하였다. 미꾸라지 apoA-I mRNA는 RT-PCR 분석을 통해 간 및 뇌 조직에서 분석한 다른 조직보다 유의적으로 높게 발현하는 것으로 나타났고, 특히 간에서 가장 높은 발현을 보였다. 수정시부터 부화 후 14일까지 초기 발생 및 치어에서의 apoA-I mRNA 발현을 조사한 결과 수정 8시간째부터 급격한 발현의 증가가 시작되어 이후 지속적으로 높은 발현 수준을 유지하였다.

Keywords

References

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