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半滑舌鳎HIRA基因部分cDNA序列克隆及表达分析
刘姗姗1,2, 张静1, 陈松林1
1.农业部海洋渔业可持续重点实验室,中国水产科学研究院黄海水产研究所,青岛 266071;2.深圳华大基因研究院, 518083
摘要:
为研究HIRA基因在半滑舌鳎胚胎发育中的作用及其组织差异表达,本实验以半滑舌鳎为研究对象,采用同源克隆策略从其精巢中分离了长度为764bp的HIRA部分cDNA序列,该片段编码254个氨基酸,具有5个WD结构域。对氨基酸进行比较发现,半滑舌鳎HIRA与比较物种的氨基酸序列具有很高的同源性,与红鳍东方鲀亲缘关系最近。运用实时定量PCR分析表明,HIRA mRNA在多细胞期到原肠胚期的表达量较高,表明HIRA对胚胎发育起重要作用;HIRA在不同组织中表达差异显著,其中在性腺中表达最高,推测HIRA在维持性腺的功能方面可能有一定作用。
关键词:  HIRA  克隆  组织差异表达  胚胎发育  半滑舌鳎
DOI:10.11758/yykxjz.20130604
分类号:
基金项目:公益性行业(农业)科研专项(200903046)、国家自然科学基金项目(31130057)、山东省泰山学者专项
HIRA gene of the half-smooth tongue-sole Cynoglossus semilaevis : cloning and its mRNA expression
Abstract:
To investigate the function of HIRA gene in the embryonic development and tissue differential expression of half-smooth tongue-sole Cynoglossus semilaevis , partial HIRA cDNA sequence of 764 bp was isolated from C. semilaevis by homologous clone method. The fragment encodes peptides of 254 amino acids, and it shares high homology with Takifugu rubripes’s HIRA gene as shown by the NJ-phylogenetic tree. It is estimated that the HIRA gene may play a vital role in embryonic development of C. semilaevis, supported by the relatively higher mRNA expressions of HIRA at early developmental stages as shown by Real-time PCR analysis. In addition, HIRA mRNA was expressed extremely high in gonads compared to the other tissues, which indicates that HIRA gene may also help gonads to maintain their normal function.
Key words:  HIRA  Clone  Tissue differential expression  Embryonic development  Half-smooth tongue-sole Cynoglossus semilaevis