黄鳝粘液雌雄差异的cDNA-SRAP分析

采用cDNA-SRAP技术对黄鳝雌雄粘液进行基因差异表达分析,筛选黄鳝体表粘液中与性别相关的分子标记。从64对引物组合中筛选出12对引物组合,共获得123个稳定清晰的扩增位点,并筛选出4条雌雄有差异的条带。这4条特异性条带经过回收、克隆、测序后,将测序结果进行Blast分析,结果发现在GenBank中只有片段1与白鳍豚硫酸盐/硫代硫酸盐CYSA样进口ATP结合盒蛋白有较高的同源性。根据片段1的序列设计1对引物进行半定量RT-PCR反应,结果发现,片段1在雌雄黄鳝体表粘液均可获得扩增条带,但其在雄性粘液中的表达量显著高于雌性。这可能是由于雌雄黄鳝粘液中所存在的表皮细胞和粘液腺细胞有差异所致。

Difference analysis of mucus between female and male ricefield eel by cDNA-SRAP

The cDNA-SRAP technique was applied to analyze the difference of gene expression in the mucus between female and male ricefield eel and to screen sex-related molecular markers.Twelve polymorphic markers selected from 64 pairs of primer were used for cDNA-SRAP amplification and 123 polymorphic loci and 4 sex-special fragments were obtained.After gel extraction,cloning and sequencing,4 sex-special sequences were obtained and submitted to db GSS (data of genome sequence survey).The result of blast analysis showed that only one sequence hit the Lipotes vexillifer sulfate/thiosulfate import ATP-binding protein CysA-like,which was used to design one pair of primer for semi-quantitative RT-PCR.PCR product was amplified in mucus of both female and male ricefield eel.However,there was a significant difference in their expression levels,with the expression in male significantly higher than in female,which may due to the difference of skin cells and mucous gland cells between female and male.