发情期湖羊卵巢组织差异表达基因的筛选与分析

Screening and analysis of differentially expressed genes of ovary tissue in Hu sheep at the estrous stage

为了筛选与湖羊发情相关的候选功能基因,试验以发情期湖羊卵巢组织cDNA为试验组,乏情期的湖羊卵巢组织cDNA为对照组,采用抑制性消减杂交（SSH）技术构建湖羊卵巢组织消减cDNA文库,并对筛选获得的功能基因进行生物信息学分析。结果表明：应用SSH技术成功构建了湖羊发情期卵巢组织消减cDNA文库;共挑取89个克隆进行测序分析,获得27个已知物种的同源基因序列和3个Novel序列;随机挑取16个阳性克隆进行PCR扩增,插入片段主要分布在150~750 bp之间;对27个已知同源基因进行功能分类,其中酶功能相关基因5个、核糖核酸结合功能相关基因10个、载体运输功能相关基因2个、调节功能相关基因3个、转运功能相关基因3个、未分类基因4个。说明试验成功筛选获得湖羊发情相关候选功能基因。

In order to screen candidate genesrelated toestrus characterin Hu sheep,the c DNA subtracted library of ovary tissue was constructed by suppression subtractive hybridization（SSH） with c DNA from ovary tissue during oestrus as experimental group and c DNA from ovary tissue during anestrus as control group. The selected functional genes were analyzed by bioinformatics.The results showed that the subtracted c DNA library of oestrous ovary tissue was successfully constructed by SSH technique.Twenty-seven sequences homologous to the genes of known species and three Novel sequences were obtained by DNA sequencing from 89 positive clones picked randomly. Sixteen positive clones were randomly selected for PCR amplification,and the inserts were mainly distributed between 150 bp and 750 bp. Twenty-seven known homologous genes were classified according to functional differences. Among them,there were 5 genes related to enzyme function,10 genes related to ribonucleic acid binding function,2 genes related to carrier transport function,3 genes related to regulatory function,3 genes related to transport function,and 4 unclassified genes.The results indicated that the candidate genes related to estrus of Hu sheep were successfully screened out in the experiment.