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vasa蛋白是DEAD-box家族蛋白的一员,在真核生物原生殖细胞形成过程中起关键作用。本实验利用RACE技术克隆获得了马氏珠母贝vasa基因(Pm-vasa),并对其结构和组织表达模式进行了分析。结果表明:Pm-vasa c DNA序列全长为1 709 bp,其中开放式阅读框1 431 bp、5’UTR 148 bp、3’UTR 131 bp,共编码476个氨基酸,分子量为52.139 k D,理论等电点为6.24。SMART软件分析显示Pm-vasa蛋白具有典型的DEAD-box结构域,且具DEAD-box家族蛋白9个典型保守基序。多序列比对结果表明Pm-vasa与紫贻贝vasa同源性最高,为74%;系统进化分析发现,Pm-vasa与紫贻贝等软体动物聚为一支。组织表达定量分析发现Pm-vasa基因m RNA在性腺中显著高表达。我们的研究结果表明Pm-vasa可能参与马氏珠母贝的性腺发育。
Vasa protein, a member of the DEAD-box family of proteins, plays a key role in the eukaryotic germ-forming process. In this study, the vasa gene (Pm-vasa) was cloned by RACE technique and its structure and tissue expression patterns were analyzed. The results showed that the full-length Pm-vasa cDNA sequence was 1 709 bp in length, with an open reading frame of 431 bp, a 148 bp 5’UTR and a 131 bp of 3 ’UTR, encoding a total of 476 amino acids with a molecular mass of 52.139 kD. The isoelectric point is 6.24. SMART software analysis showed that the Pm-vasa protein has a typical DEAD-box domain with 9 typical conserved motifs of the DEAD-box family of proteins. The results of multiple sequence alignment showed that Pm-vasa had the highest homology with Vasa vizslavii, accounting for 74%. Phylogenetic analysis showed that molluscs such as Pm-vasa and Mytilus coruscus were clustered together. Tissue expression quantitative analysis found that Pm-vasa gene m RNA was significantly high expression in the gonads. Our results suggest that Pm-vasa may participate in gonadal development in Pinctada martensii.