梅山猪与杜洛克猪中等卵泡差异表达基因的鉴定

Identification of Differentially Expressed Genes in Medium-sized Ovarian Follicles between Meishan and Duroc Sows

本试验采用抑制性消减杂交技术构建了卵泡期第4天梅山猪和杜洛克猪中等卵泡M2组织差异表达的消减cDNA文库,从中筛选差异表达的基因,并通过实时荧光定量PCR对其进行验证,利用DAVID软件对差异表达的基因进行聚类分析。结果表明,从梅山猪和杜洛克猪M2卵泡消减文库中筛选得到了148和75个差异表达的ESTs,分别含有125和60个已知的基因。实时荧光定量PCR验证结果与筛选结果相符。GO功能分类注释到调控代谢、细胞循环、生物合成、胞内转运、类固醇雌激素受体和刺激等生物学过程。KEGG Pathway分析表明有6个基因参与TGF-beta信号通路,5个基因参与卵母细胞减数分裂信号通路,7个基因参与类固醇雌激素受体信号通路,推测TGF-beta信号通路中的基因可能调控猪卵泡的发育。研究结果为深入探讨卵泡发育机理和对繁殖性状影响的遗传机理奠定了基础。

To reveal the molecular mechanism involved in different number of ovulation between hyperprolific and ordinary sows,forward and reverse subtracted cDNA libraries were constructed to screen differentially expressed genes in medium ovarian follicles at 4th day during follicular phase of estrous cycle between Meishan and Duroc sows.The differentially expressed genes selected were demonstrated by Real-time PCR and cluster analysis was performed through DAVID.The results showed that a total of 148 and 75non-redundant ESTs were isolated from the SSH library of Meishan and Duroc sows M2 follicles,including 125 and 60known genes,respectively.The results of Real-time PCR were consistent with the screening results.GO analysis indicated that these genes were involved in regulation of metabolic process,cell cycle,biosynthetic process,intracellular transport,steroid hormone receptor and stimulus.KEGG pathway analysis defined 6genes involved in TGF-beta signaling pathway,5genes in oocyte meiosis pathway,7genes in steroid hormone receptor signaling pathway.Genes controlling TGF-beta signaling pathway were considered to play an important role in regulating follicle development.The research would be helpful for further studying the follicular development and the genetic mechanism on re-productive traits.