马氏珠母贝Pm-vasa基因的克隆与表达分析  

Cloning and Expression Analysis of vasa Gene from Pinctada fucata martensii

作　　者：刘雅[1,2] 王庆恒[1,2] 黄荣莲[1,2] 焦钰[1,2] 杜晓东[1,2] 

机构地区：[1]广东海洋大学水产学院,湛江524088 [2]广东省珍珠养殖与加工工程技术研究中心,湛江524088

出　　处：《基因组学与应用生物学》2017年第7期2832-2839,共8页Genomics and Applied Biology

基　　金：广东省海洋渔业与产业发展专项(Z2014009);国家自然科学基金(31372526);广东海洋大学创新强校工程(GDOU2014050207)共同资助

摘　　要：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 is a member of the DEAD-box family proteins and plays a key role in the formation of primordial germ ceils in eukaryotes. We cloned the full length ofPm-vasa cDNA by rapid amplification of cDNA end （RACE） methods and analyzed its structure and expression pattern. The results showed that： the full length of Pm-vasa cDNA was 1 709 bp, containing an open reading frame （ORF） of 1 431 bp, a 5＇UTR of 148 bp and a YUTR of 131 bp, it encoded 476 amino acids. The predicted molecular weight was 52.139 kD, and the isoelectric point was 6.24. SMART sofLware analysis showed that protein Pm-vasa had a typical DEAD-box domain and contained nine conserved motifs of the DEAD-box family. Multiple sequence alignment results showed that high identity of vasa in amino acid sequence （more than 74%） was observed between P. Fucata martensii and Mytilus galloprovincialis. Phylogenetic analysis showed that Pm-vasa was in the same cluster with Mytilus galloprovincialis and other molluscs. Using real-time PCR technology, we found that Pm-vasa was highly expressed in the gonads. Our results indicated that Pm-vasa might take part in the gonad development ofP. fucota rnartensii.

关 键 词：马氏珠母贝 Pm-vasa基因 基因克隆 表达分析 

分 类 号：Q78[生物学—分子生物学] S917.4[农业科学—水产科学]