抑制消减杂交技术筛选鹅产蛋期差异表达基因

Differentially Expressed Geese Genes in Egg Laying Period by Using SSH

[目的]鉴定筛选鹅产蛋与就巢差异表达基因。[方法]应用抑制消减杂交技术构建鹅产蛋与就巢鹅卵巢组织双向cDNA文库,获得差异表达基因。[结果]从双向文库随机挑选单菌落进行PCR验证,结果表明文库质量良好。选取654个阳性克隆进行测序分析,获得641条表达序列标签(ESTs)。对ESTs进行去除载体序列、质量检测、聚类及拼接,经Blast比对,有166条ESTs找到与之匹配的同源序列,其中92个是功能基因。[结论]比较就巢与产蛋SSH文库,发现产蛋特异性基因文库中参与物质能量代谢、细胞防御的基因较多,而在就巢SSH库中参与细胞凋亡、信号转导及细胞结构的基因出现频率较高。该结果对进一步研究鹅卵泡发育及繁殖力分子标记筛选打下了基础。

[Objective]The aim was to identify and screen differential expression genes between egg-laying and broodiness phase.[Method]SSH was applied to construct directional cDNA libraries of geese egg-laying and broodiness ovarian tissue,thus,differential expression genes were obtained.[Result] Positive clones from libraries were randomly selected to validate with PCR,the results showed that the quality of libraries were perfect.Consequently,654 positive clones were selected to sequence,qualify,cluster,assemble and comparing through Blast.166 differentially expressed gene fragments from two libraries showed homology to other known sequences,including 92 functional genes.[Conclusion]Comparing broodiness library with SSH library,it indicated that some laying-specified genes involved in metabolism and cell defense;on the other hand,some broodiness-specified genes involved in apoptosis,cell organization,signal transduction.The results provided the basis for future revealing the mechanism on ovarian follicles development and molecular marker of fertility.