凡纳滨对虾Rab6A基因的克隆及表达  

Cloning and Expression of Rab6A Gene From Litopenaeus vannamei

作　　者：岳磊磊[1] 邵朋威[1] 宗自杰[1] 赵永贞[2] 陈晓汉[2] 郑喜邦[1] 陈秀荔[2] 

机构地区：[1]广西大学动物科技学院,广西南宁530005 [2]广西水产研究所广西水产遗传育种与健康养殖重点实验室,广西南宁530021

出　　处：《动物医学进展》2012年第8期22-26,共5页Progress In Veterinary Medicine

基　　金：广西自治区直属公益性研究专项(2060302CXIF-2009-02);现代农业产业技术体系建设专项(nycytx-46);重点实验室开放基金(GXKL-AQUA-2011-03)

摘　　要：为研究凡纳滨对虾免疫调节因子Rab蛋白在对虾机体的免疫调节过程中的作用机理,克隆了凡纳滨对虾Rab6A基因,并对其在感染对虾传染性皮下及造血组织坏死病毒(IHHNV)对虾不同组织中的表达情况进行了研究。根据日本对虾Rab蛋白基因设计引物,采用RT-PCR方法克隆了长为558bp的Rab6A基因的部分cDNA序列,其开放阅读框为442bp,可编码147个氨基酸,推算其分子质量约为16.977ku,理论等电点为4.828,Rab6A与其他物种的Rab基因比对发现,该基因氨基酸序列具有较高的保守性。半定量PCR结果显示,Rab基因在不同组织中的表达情况没有明显的组织特异性,肝胰腺中表达量比其他组织略高,而在感染IHHNV的对虾心脏、肝胰腺、肠道、胃、腮、肌肉、血液组织中比正常组织中表达量增加,表明Rab蛋白参与抗病毒免疫反应。To study the molecular mechanism of Litopenaeus vannamei Rab protein in immune regulation, the re- search cloned Rab6A gene of Litopenaeus vannamei, and studied its expression character in different tissues of in- fected Infectious hypodermal＆ haematopoietic necrosis virus（IHHNV） shrimp and uninfected-IHNV shrimp. Based on the sequence from Marsupenaeus japonicus, a pair of primers were designed, and a 558bp cDNA was ob- tained by RT-PCR. The cDNA contained 442 bp ORF encoding a peptide of 147aa. The predicted molecular weight was about 16. 977 ku, and its calculated isoelectric point（pI） was 4. 828 . Rab6A was compared with other species through BLAST analysis, indicating that the amino acid sequence of Rab6A gene wsa highly conserved. Tissue expression pattern by semi-quantitative PCR revealed that Rab6A was not evident tissue specificity, the ex- pression of the gene in hepatopancreas was higher than that in other tissues, while the expression of the gene in myocardium, hepatopancreas, intestinal, gastric,gill, muscle, blood of infected IHHNV shrimp were higher than that of uninfected IHHNV shrimp. The result indicated that Rab6A protein involved antiviral immune reaction of Litopenaeus vannamei.

关 键 词：凡纳滨对虾 传染性皮下及造血组织坏死病毒 Rab基因 免疫反应 

分 类 号：Q785[生物学—分子生物学] S945.19[农业科学—水产养殖]