[논문]Isolation and Molecular Phylogeny of Three Muscle Actin Isoforms of an Endangered Freshwater Fish Species Hemibarbus mylodon (Cypriniformes; Cyprinidae)

김근용; 남윤권

[국내논문] Isolation and Molecular Phylogeny of Three Muscle Actin Isoforms of an Endangered Freshwater Fish Species Hemibarbus mylodon (Cypriniformes; Cyprinidae) 원문보기

韓國養殖學會誌 = Journal of aquaculture, v.22 no.1, 2009년, pp.83 - 91

김근용 (Department of Aquaculture, Pukyong National University) , 남윤권 (Department of Aquaculture, Pukyong National University)

Abstract ▼ AI-Helper

The Korean doty barbel Hemibarbus mylodon (Cypriniformes; Cyprinidae) is a critically endangered freshwater fish species mainly because of its natural habitat degradation. Three full-length complementary DNA (cDNA) clones representing different muscle actin isoforms were isolated and characterized. The three muscle actin isoforms were 1,294-1,601 bp long with the identical open reading frames of 1,134 bp with the deduced amino acid residues of 377. They showed 83.9-87.2% identities in the coding nucleotide level and 96.8-98.1% identities in the amino acid level. Phylogenetic analysis with the coding nucleotide sequences revealed that three muscle actin isoforms of H. mylodon formed strongly supported monophyletic groups with one of cypriniform skeletal $\alpha$-actin (acta1), cypriniform aortic $\alpha$-actins (acta2), and uncharacterized Danio rerio muscle actin isoform/Salmo trutta slow muscle actin (a novel muscle actin type). Our phylogenetic tree further suggested that cypriniform acta2 only showed the orthologous relationship to tetrapod acta2. Other multiple actin isoforms from diverse teleostean taxa were however clustered to no tetrapod orthologs, i.e., acta1, cardiac $\alpha$-actins (aetc1), acta2, and enteric $\gamma$-actin (actg2). This result strongly suggested that teleostean muscle actins have experienced different and complicated evolutionary history in comparison to mammalian counterparts.

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제안 방법

, 2007). In this study, we isolated three muscle actin mRNA species from H. mylodon and carried out the phylogenetic analysis within the vertebrate lineage to reveal their orthologous rela tionships and also to fiirther discuss evolutionary history of teleost muscle actins in comparison to tetrapod counterparts.
RT-PCR product was cloned into pGEM®-T Easy Vector (Promega, Madison, WI, USA) according to the manufacturer's instruction. The insert DNA was read at both directions using ABI 3700 Automatic Sequence Analyzer (Applied Biosystems, Foster City, CA, USA) in order to confirm the sequence of each isoform. The muscle actin isoforms isolated in this study were deposited in GenBank under accession numbers, FJ713567-FJ713569.
trutta slow muscle actin, which were designated as novel muscle actin type 1 in this study. Given the presence of as much as seven muscle actin isoforms in D. rerio (Fig. 3) we would expect the pres ence of additional muscle actin isoforms in H. mylodon when extending EST sampling.

대상 데이터

Fish specimens used in this study were adult individuals maintained at Soonchunhyang University, Korea. Informa tion on the H.

이론/모형

The nucleotide sequence matrix was partitioned into three codon positions for coding nucleotide sequences. Model selection strategy of Akaike Information Criterion (AIC) implemented in MrModeltest 2.2 (Nylander, 2004) was used to determine the best-fit evolutionary model of nucleotide substitutions for each partition, and the GTR+I+r (Iset nst = 6 rates = invgamma) was selected for all three positions. All model parameters were unlinked [unlink statefreq = (all) revmat (all) 아i햐泪 = (all) pinvar = (all)], and all partitions were allowed to have differ ent rates (prset ratepr = variable).

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