兰州鲇MyoD基因的克隆与生物信息学分析

为研究兰州鲇(Silurus lanzhouensis)生长性状相关基因,运用RT-PCR、TA克隆和核酸测序等技术对兰州鲇MyoD基因的CDS区及全基因进行克隆和生物信息学分析。获得兰州鲇MyoD基因的完整CDS区序列810 bp(Gen Bank登录号:KT277551)及MyoD基因全序列1 210 bp(Gen Bank登录号:KT339175),包括部分5'端63 bp和3'端58 bp;ORF区含有3个外显子和2个内含子,外显子长度分别为510 bp、80 bp和220 bp,内含子长度分别为156 bp和123 bp,编码269个氨基酸残基组成的可溶酸性蛋白质;预测亚细胞定位MyoD主要分布于细胞核(56.5%),MyoD二聚体结构是一个螺旋-环-螺旋(b HLH)。基于12种鱼类MyoD基因CDS序列构建系统发育树及编码区同源性比较,分析结果表明,MyoD基因编码区在进化过程中比较保守,且兰州鲇MyoD与斑点叉尾、白鲶鱼、蓝鲶鱼之间存在较高的同源性。

Cloning and Bioinformatics Analysis of MyoD Gene in Silurus lanzhouensis

In order to explore the gene related to growth trait of Silurus lanzhouensis, MyoD gene of S.lanzhouensis was cloned and analyzed with RT-PCR , TA cloning and nucleic acid sequencing technology. The810 bp length CDS region (GenBank accession No.: KT277551) and 1 210 bp length whole gene(GenBank accession No.: KT339175) of MyoD was obtained, the 1 210 bp length whole gene included 5′UTR in 63 bp length and 3′UTR in 58 bp length, and ORF in-cluded three exons and two introns , the length of three exons was 510 bp, 80 bp and 220 bp, and the length of two introns was 156 bp and 123 bp.A total 269 amino acids were encoded by the MyoD gene.Subcellular localization of MyoD was mainly in nucleus (56.5%).The secondary structure of MyoD was a helix loop helix (bHLH).Phylogenetic tree and the Homology of twelve fish based on MyoD gene coding region analysis showed that the encoding of MyoD gene sequences were highly conservative, S.lanzhouensis had a higher homology with Ictalurus punctatus, Ameiurus catus and Ictalurus furcatus.