施氏鲟精巢和卵巢差减cDNA文库的构建和差异表达EST的筛选  

Cloning and screening of differentially expressed genes from testis and ovary of Amur sturgeon

作　　者：陈金平[1] 王斌[2] 吴文化[2] 梁冰[1] 张树义[1] 

机构地区：[1]中国科学院动物研究所,北京100080 [2]中国水产科学研究院鲟鱼繁育技术工程中心,北京100039

出　　处：《中国水产科学》2006年第1期8-12,共5页Journal of Fishery Sciences of China

基　　金：国家自然科学基金杰出青年基金项目(30025007);中国科学院知识创新工程重要创新方向项目(KSCX2-SW-118)

摘　　要：应用抑制差减杂交(SSH)技术构建了施氏鲟(Acipenser schrenckii)精巢和卵巢组织的SMART cDNA文库及其cD-NA差减文库,从两个差减文库中随机挑取200个克隆进行PCR检测,随机挑选其中的123个cDNA克隆测序。将所测序列经GenBank检索和生物信息学比较,发现有55个cDNA片段序列在GenBank上无明显的同源性,68个片段与已报道的基因有较高的同源性,其中17个cDNA片断为精巢中特异表达的基因,而51个为卵巢中特异表达的基因。精巢中大量表达的主要为延长因子(EF-1)和脂肪酸连接蛋白等与精子生成发育有关的基因,而卵巢主要为ZPC、组蛋白和周期蛋白等与卵细胞发育成熟相关的基因。这些EST数据为进一步筛选和克隆鲟鱼性腺组织特异性表达基因提供了材料平台,为鲟鱼基因组数据增补了大量信息。[中国水产科学,2006,13(1):8-12]The Amur sturgeon （Acipenser schrenckii ） is an ancient freshwater Chondrostei species, which is mainly distributed in Asia, including China. This species represents an economically important source of food including caviar. However, little genetic information is available for Amur sturgeon, and there are currently no genetic markers for selection of broodstock or manipulation of its sex. Identification of sex related genes in Amur sturgeon provides data for studying evolution and function and the developmental biology of Amur sturgeon. To find these sex development related genes, A systematic study was initiated to identify differentially expressed genes in the testis and ovary of the Amur sturgeon by using suppression subtractive hybridization （SSH） technique. Some differentially expressed genes between female gonad （FG） and male gonad （MG） in the Amur sturgeon were obtained. Two SSH plasmid libraries for gonads of Amur sturgeon were constructed. And then differentially expressed genes were sequenced. Searching GenBank by using these nucleotide sequences indicated that 55 cDNA fragments could not find their homologous sequences in the database. Of the 68 known genes, 17 cDNAs were differentially expressed in testis, and were made up of fatty acid-binding protein, Collagen type I alpha 2, EF-1, et al. , and 51 cDNAs were differentially expressed in ovary, which were mainly ZPC, Histone protein（H2A, H4,H3）, Cyclin, et al., closely relevant to egg development whose expression may be good for producing good quality caviar. The results are good enough for further cloning of the gonads specific expression genes and adds much genetics information of sturgeon.

关 键 词：施氏鲟 差减CDNA文库 差异表达 抑制差减杂交 

分 类 号：Q959.463[生物学—动物学]