脊尾白虾(Exopalaemon carinicauda)vasa基因cDNA克隆及其在卵巢发育中的表达分析  

CLONING AND EXPRESSION OF GENE VASA DURING OVARIAN DEVELOPMENT CYCLE IN EXOPALAEMON CARINICAUDA

作　　者：徐文斐 刘萍[2] 李吉涛[2] 李健[2] 陈萍[2] 

机构地区：[1]大连海洋大学水产与生命学院,大连116000 [2]中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《海洋与湖沼》2014年第3期136-144,574,共9页Oceanologia Et Limnologia Sinica

基　　金：国家高技术研究发展计划课题(2012AA10A409);山东省自主创新专项(2013CX80202);国家虾产业技术体系(CARS-47);公益性行业(农业)科研专项(201103034);中国水产科学研究院院级基本科研业务费专项资金资助项目(2013A0701)

摘　　要：VASA蛋白属于DEAD-box家族蛋白,是一种RNA解旋酶,在生殖细胞的分化中具有重要作用。本研究从脊尾白虾(Exopalaemon carinicauda)卵巢中克隆得到vasa基因cDNA全长(将其命名为Ec-vasa),Ec-vasa序列长度为2516bp,开放阅读框(ORF)长度为1800bp,编码600个氨基酸,具有DEAD-box家族蛋白的8个高度保守的结构域,与日本沼虾(Macrobrachium nipponense)的同源性最高(80%)。半定量RT-PCR结果显示,Ec-vasa在卵巢组织中特异表达,在肌肉、肝胰腺、心脏和鳃等组织中均未检测到表达。荧光定量结果显示,Ec-vasa在卵巢发育Ⅰ期具有较高表达量,随着卵巢的发育,表达量逐渐降低,在第Ⅳ期达到最低水平。结果表明,Ec-vasa基因可能在脊尾白虾卵巢的早期分化和卵子发生过程中起重要作用。The vasa gene encodes DEAD box family protein of ATP-dependent RNA helicase,and it plays a crucial role in primordial germ cell proliferation.In this report,a vasa gene was isolated from Exopalaemon and named Ec-vasal carinicauda using reverse transcription polymerase chain reaction(RT-PCR) coupled with rapid amplification of cDNA ends(RACE) approaches.The Ec-vasa cDNA sequence comprises 2516bp with an open reading frame of 1 800bp encoding 600 amino acids.The deduced amino acid sequence contains 8 conserved motifs of DEAD-box family protein.Comparisons in the deduced amino acid sequence with those of other invertebrates revealed the highest homology(80%) with Macrobrachium nipponense.In the organs of the adult,the Ec-vasa transcripts were detected in ovary.In the ovarian development cycle,the highest mRNA expression level of vasa gene was found instageⅠ,and then the transcripts decreased with ovarian development.As the time progressed,the expression level of Ec-vasa reached the lowest level at stage Ⅳ.These results indicate that Ec-vasa gene may play an important role in oogenesis during ovarian development cycle,and it was expected as an effective molecular marker to the origin of PGCs and gonadogenesis.

关 键 词：脊尾白虾 卵巢发育 表达量 日本沼虾 开放阅读框 基因片段 卵巢组织 克隆 生殖细胞 卵母细胞 

分 类 号：P7[天文地球—海洋科学] P9