山羊高繁殖力相关基因的筛选与分析

Screening and Analyzing of Genes on Prolificacy in Goat

为查找与山羊繁殖力相关的基因,进一步研究繁殖力调控的遗传机制,本研究选用12只冀中山羊,按照其繁殖记录分为高产和低产2组,应用差异显示反转录PCR(DDRT-PCR)技术,分析了山羊在发情期卵巢组织基因表达的差异。经二次扩增、反式Northern以及半定量PCR验证分析,最终得到4个表达量差异的阳性片段。所得片段经克隆测序提交至GenBank,并进行BLAST分析。结果,4个差异片段中2条为新发现的表达片段,在NCBI中未发现高度同源序列。1条与野猪中获得的克隆片段THY010003E05同源性为81%,但功能未知。另1条差异片段的序列与牛PCNP的mRNA序列的同源性为94%。结果提示,该片段所属基因应属于PCNP基因,结合对其功能的分析结果,推测PCNP可能通过参与卵细胞生成过程中的细胞周期调控影响山羊繁殖力。

To search the genes associated with prolificacy of goats,and research the genetic mechanism of fertility regulation,twelve Jizhong goats were divided into 2 groups according to their reproductive records.Differential display reverse transcript PCR（DDRT-PCR） method was used in the analysis of the discrepancy of gene expression in ovarian tissue of goats in estrual period.4 segments differentially expressed in quantity were finally obtained via second PCR amplification,reverse Northern blot and semi-quantitative PCR.After sequencing and cloning,these segments were submitted to GenBank,and analysised by BLAST.Two of the 4 were new segments and high homologous sequences in NCBI was not found.Another segment shared 81% similarity with one clone segment（THY010003E05） of wild boar（Sus scrofa）,but with unknown function.The other segment was 94% similar to the mRNA of PCNP gene in cattle.It indicate that the segment is a homolog of PCNP gene and it could affect goat prolificacy via involving in cell cycle regulation of oocyte according to the analysis of its function.