方格星虫dmrt1基因的克隆和表达分析

CLONING AND EXPRESSION ANALYSIS OF PEANUT WORMS TRANSCRIPTION FACTOR DMRT1

为了研究dmrt1(Double sex and mab-3 related transcription factor 1)基因在方格星虫(Sipunculus nudus)的表达特征,首先采用RACE方法克隆了方格星虫dmrt1基因。方格星虫dmrt1的cDNA全长1509 bp,开放阅读框615 bp,共编码204个氨基酸。生物信息学分析表明方格星虫DMRT1的DM结构域具有一定的保守性,与其他物种的同源性能够达到70%以上。成体组织荧光定量PCR分析表明方格星虫dmrt1仅在方格星虫雄性个体的体腔液中表达,通过原位杂交显示dmrt1定位于体腔液精子团周围的滋养细胞。这些结果提示,方格星虫dmrt1可能在调控方格星虫的雄性个体的生殖发育过程中发挥着重要作用。

The DMRT1(double sex and mab-3 related transcription factor 1)is a highly conserved transcriptional regulator to play important role in sex determination and differentiation in most species ranging from nematodes to mammals.In this study,we cloned the cDNA sequence of dmrt1 from peanut worms cDNA library by RACE-PCRs.The full-length dmrt1 cDNA was 1509 bp with a 615 bp open reading frame(ORF)encoding a protein of 204 amino acids.Bioinformatics analysis showed that peanut worms DM domain shared high homology(>70%)with those of other vertebrates as well as the invertebrate marine mollusks DMRT1.dmrt1 was abundantly expressed in coelomic fluid of male peanut worms,where existed the germ cells.In situ hybridization explored positive dmrt1 signals in the trophoblast cells.These results suggested that dmrt1 may play important role in male germ cell development,and reproductive regulation in spermatocyte maturation of peanut worms.