文蛤HDAC1基因克隆、时空表达及生长相关SNP位点筛查

CLONING, SPATIOTEMPORAL EXPRESSION AND SNPS IDENTIFICATION OF HDAC1 GENE IN HARD CLAM MERETRIX MERETRIX

为探索HDAC1基因在文蛤生长发育中的作用,研究通过已构建的文蛤转录组文库,利用SMART RACE技术扩增得到文蛤HDAC1(Mm-HDAC1)基因的c DNA全长序列,分析了其生物信息学、组织及发育阶段表达特征,并用直接测序法在外显子中筛查生长相关的SNP位点。结果显示,Mm-HDAC1基因的c DNA全长3065 bp,开放阅读框1599 bp,编码532个氨基酸;氨基酸序列比对发现,文蛤与其他物种同源性为74.3%—78.7%。荧光定量PCR(q RT-PCR)结果表明,Mm-HDAC1基因在文蛤6个组织中均有表达,其中外套膜中的表达量相对最高,并与其他组织间有极显著差异(P<0.01);不同发育时期的表达差异结果表明,MmHDAC1基因在壳顶幼虫期表达量最高,显著高于其他发育时期(P<0.05)。SNP位点筛查结果表明,在MmHDAC1基因的外显子区域发现了19个SNP位点,其中有3个SNP位点(627A>T、924T>C和1266T>C)与文蛤生长相关。

Mm-HDAC1 cDNA was cloned by SMART RACE techniques using 454 cDNA library ofMeretrix meretrix. The bioinformatics and expression profiles in different tissues and developmental stages were analyzed, and SNPs were screened. The results indicated that the full length cDNA ofMm-HDAC1 gene was 3065 bp, containing a complete 1599 bp ORF that encodes 532 amino acids. Comparison of animal acid sequence, 34. meretrix shared 74.3%-78.5% identity with others, suggesting that Mm-HDAC1 was highly conservative. The result of qRT-PCR showed that MmHDAC1 expressed in all six tissues, and its expression was significantly higher in mantle than other tissues （P〈0.01）. Moreover, the expression of Mm-HDAC1 gradually increased with the process of the development with the highest level in umbo larvae stage （P〈0.01）. 19 SNPs in exons of Mm-HDAC1 were identified and 3 SNPs were potentially associated with M. meretrix growth （627A〉T, 924T〉C and 1266T〉C）.