奥利亚罗非鱼和尼罗罗非鱼SRY基因同源克隆和序列分析  

Cloning and Analysis of SRY Homologous Sequences from Oreochromis aureus and O. nilotica

作　　者：杨弘[1] 夏德全[1] 许广平[1] 何杰[1] 曹丽萍[1] 吴婷婷[1] 

机构地区：[1]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,无锡214081

出　　处：《农业生物技术学报》2005年第4期524-527,共4页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2004AA243060);国家自然科学基金(No.30371116);无锡市自然科学基金项目(No.CK030001);中国水产科学研究院人才资金A类资金项目资助。

摘　　要：采用人SRY基因保守序列的特异性引物,对奥利亚罗非鱼(Oreochromisaureus)和尼罗罗非鱼(O.nilotica)雌、雄个体基因组进行PCR扩增,回收其中的600bp片段,分离克隆并双向测序。用DNAStar分析比较所得序列,表明克隆的600bp片段为SRY基因的同源序列。对两种罗非鱼雌、雄个体SRY同源序列进行多序列比较分析(CLUSTALW分析),表明罗非鱼间SRY同源序列的相似性非常高,但仍存在性别间、种间差异。奥利亚罗非鱼雌、雄间序列相似性为99.5%,尼罗罗非鱼雌、雄间为99.5%,奥利亚罗非鱼与尼罗罗非鱼种间则为98.7%,罗非鱼与人类SRY基因间只有7.1%,结果为进一步研究罗非鱼SRY同源基因与性别决定的关系提供了可能性。根据这些结果构建了罗非鱼系统分类树,表明奥利亚罗非鱼与尼罗罗非鱼具有很近的亲缘关系。Using the primers which specially amplify the conservative motif of human SRY gene, the PCR amplification of SRY homologous sequences in genomic DNA from male and female individual of Oreochromis aureus and O. nilotica was studied. The 600 bp fragments of PCR amplification were isolated and cloned in plasmid vector pUCm-T. By sequencing and DNAStar software analysis, the result showed the 600 bp fragments were human SRY homologous sequences. Multiple sequence alignments by CLUSTAL W indicated that there was high genetic similarity between the four SRY-related sequences of tilapia. The percent identity of SRY-related sequence between male and female O. aureus was 99.5%, the percent identity between male and female O. nilotica was 99.5%, and that between O. aureus and O. nilotica was 98.7%. The result made it possible to study the correlativity between SRY homologous gene and sex-determing oftilapia. And then phylogenetic tree of O. aureus and O. nilotica was constructed, which indicated that their relationships were close.

关 键 词：奥利亚罗非鱼 尼罗罗非鱼 SRY基因 同源克隆 基因序列 亲缘关系 

分 类 号：Q959.46[生物学—动物学]