猪GnRH受体基因多态性分析及相关基因组织表达谱的构建

促性腺激素释放激素(GnRH)受体基因与猪的繁殖性状密切相关,本试验对不同猪种的GnRH受体基因的编码区进行了SSCP分析,未发现多态性。在GnRH受体基因5′侧翼序列上设计引物扩增了该基因大约2000 bp的侧翼序列,通过比对设计引物,对其中的一段含有GAGABox的插入突变序列和一段由于点突变而形成转录调控蛋白结合序列IRF-1的区域运用SSCP的方法进行了多态性分析。结果表明在插入突变处,基因频率在不同的猪种间存在着很大的差异性,其中民猪与其他猪种的差异性最大。IRF突变只存在于民猪中,并且只存在AA型和AB型。构建了3头大白猪母猪GnRH基因、GnRH受体基因、卵泡抑素(FST)基因、促卵泡素(FSH)基因、FSH受体基因、促黄体素(LH)基因、抑制素(inhibin)β(a)基因、inhibinβ(b)基因的组织表达谱,根据表达谱可以推测在下丘脑-脑垂体-性腺轴外不存在这些基因表达产物的相互作用系统。

Polymorphism analysis on the GnRH receptor gene and tissue-expression patterns of related genes in the pig

The GnRH receptor gene is closely related to reproduction performance.The SSCP method was carried on the coding region of GnRH receptor gene in varied breeds of pigs and no polymorphism was found in the current project.About 2 000 bp was amplified by the primers designed on the 5′flanking sequence of GnRH receptor gene.By comparing two pairs of primers designed by the SSCP method and two mutation sequences,one was an insert mutation within a GAGA Box,the other was a single nucleotide mutation creating a transcription factor IRF-1 binding sit.The analysis results indicated that the genotype frequency had a great discrepancy in the insert mutation varying according to the boar species.The min-pig had a very significant discrepancy compared with other breeds of pigs.The IRF-1 mutation had only in the min-pig.Expression spectrums of GnRH,GnRHR,FST,FSH,FSHR,LH,inhibin β(a) and inhibin β(b) were constructed with three female large white pigs.According to the expression patterns,it suggested that there were no outside hypothalamus-pituitary-gonad axis interaction system of these genes.