用链球菌疫苗免疫罗非鱼前后其差异表达基因的鉴定与分析  

Identification and analysis of differently expressed genes of tilapia immuned by Streptococcus iniae vaccine

作　　者：李超[1,2] 陈明[1,3] 李莉萍[1] 黄维义[2] 余晓丽[1] 黄婷[1] 张彬[1] 雷爱莹[1] 陈福艳[1] 梁万文[1] 

机构地区：[1]广西水产研究所,广西南宁530021 [2]广西大学动物科学技术学院,广西南宁530005 [3]中国水产科学研究院淡水渔业研究中心农业部水生动物遗传育种和养殖生物学重点开放实验室,江苏无锡214081

出　　处：《大连水产学院学报》2009年第5期379-386,共8页Journal of Dalian Fisheries University

基　　金：国家科技支撑计划项目(2008BADB9B04);农业部水生动物遗传育种和养殖生物学重点开放实验室开放基金资助项目(BZ2007-05);广西科技计划应用基础研究项目(桂科基0832029)

摘　　要：采用抑制性差减杂交技术(SSH)成功构建了罗非鱼O reochrom is nilotica被链球菌Streptococcus iniae疫苗免疫前后正、反两个淋巴细胞cDNA差减文库,富集了免疫过程中表达谱发生变化的淋巴细胞基因,并对文库进行了斑点杂交筛选、测序和ESTs初步分析。结果表明:文库富集了高丰度的差异表达基因,阳性率较高。正向文库中筛选得到21个免疫期间表达丰度上调的基因,其中1个基因与罗非鱼已知基因具有相似性,10个基因与其他鱼类已知基因相似,1个基因与其他物种已知基因具有相似性,9个为未知新基因;负向文库中筛选得到24个免疫期间表达丰度下调的基因,其中3个与罗非鱼已知基因具有相似性,15个与其他鱼类已知基因相似,6个为未知新基因。进一步功能注解发现,正向文库功能基因包括免疫信号传导相关蛋白、防御相关蛋白、肿瘤免疫相关蛋白和淋巴细胞相关蛋白,这些基因免疫后表达丰度升高,提高了机体免疫抵抗力。负向文库功能基因包括细胞免疫相关蛋白、感染相关蛋白和肿瘤发生相关蛋白,这些基因免疫后表达丰度降低,可以降低机体感染和发病几率。The forward and reverse two subtractive cDNA libraries were constructed from the peripheral blood leucocytes of Nile tilapia Oreochromis nilotica vaccinated by bacterium Streptococcus iniae vaccine by i. p to screen differently expressed immune related genes to investigate regulation of immune - related gene expression by suppression subtractive hybridization（SSH） technique. The lymphocyte genes that changed in the immune process were enriched and the library had been screened by dot - blot while the ESTs were analyzed primarily. It was found that the library enriched the high - abundance differentially expressed genes, with a high positive rate. Total of 21 genes, upregulated in the forward library, were identified,in which 1 genes showed homology with known genes of genes had homology with known genes of other fish, and 1 genes showed homology with known genes of other animal,and 9 genes were unknown genes. While 24 genes, downregulated in the reverse library, were identified, in which 3 genes showed homology with known genes of 15 genes showed homology with known genes of other fish, and 6 genes were unknown genes. Further gene function analysis revealed that the forward library included immunization signal transduction related protein, disease defence related protein, tumor immunity related protein and leucocytes related protein. These genes up -regulated after immune, increased the body immune resistance. The tumorigenesis related protein, disease related protein and cell immunity related protein were found in the reverse library. These genes down - regulated after immune, and reduced the incidence of infection as the further bioinformatics analysis of this library and using correlation technology to obtain full - length secquence of immune related genes.

关 键 词：罗非鱼 链球菌 疫苗 差减文库 免疫相关基因 斑点杂交筛选 

分 类 号：Q786[生物学—分子生物学]