翘嘴红鲌Vasa基因cDNA克隆及其组织表达特征

Cloning of Vasa and Expression Analysis between Different Tissues in Erythroculter ilishaeformis

为了研究Vasa基因在翘嘴红鲌生殖细胞发育中的作用,本研究以翘嘴红鲌冬片鱼种的精巢组织为材料,提取总RNA,采用RT-PCR和RACE技术分离、克隆Vasa基因的cDNA全序列,与其他21个物种的Vasa氨基酸序列进行同源性比较,并利用荧光定量PCR分析Vasa基因在翘嘴红鲌各组织的表达。结果表明:该基因序列全长为2 713 bp,编码684个氨基酸,具备DEAD-box家族特有的8个保守基序;翘嘴红鲌与草鱼的同源性最高达96%;Vasa仅在翘嘴红鲌精巢和卵巢中表达;与成鱼繁殖期的性腺组织相比,后者表达量高,差异显著(P<0.05),同期精巢、卵巢Vasa基因表达量差异不显著。由此可见,Vasa可作为翘嘴红鲌生殖细胞的标志基因。该基因全序列的获得为今后进一步探讨翘嘴红鲌生殖发育规律奠定了基础。

In order to study the Vasa gene i n the role of Erythroculter ilishaeformis germ cell differentiation and development, we cloned and characterized the cDNA of Vasa through RACE technology. Vasa expression were investigated in different tissues in winter pieces fry and the gonad in immaturity and maturity of Erythroculter ilishaeformis using real-time quantitative PCR respectively. The size of Vasa gene is 2712 bp, which encodes a protein of 684 amino acids.The deduced amino acid sequence contains 8 conserved motifs of DEAD-box family protein. Compared with 21 other species＇ same gene sequence, it has the highest homology with grass carp （up to 96%）.The result of real-time quantitative PCR analysis showed that Vasa was only expressed in gonad, and maturity was higher than immaturity （P〈0.05）. The results suggest that Vasa gene is a biomarker in the process of germ cell development in Erythroculter ilishaeformis, which laid the foundation for future germ cell studies.