【摘要】 在梅山猪保种群和法系大白猪群体中采用直接测序法和PCR-RFLP法对基因VEGFA、HIF1A和EPAS1进行了基因多态位点的寻找及多态性检测并对不同基因型的总产仔数、产活仔数等繁殖性状进行了相关分析。在所获得的基因VEGFA的扩增片段中共检测到17个多态位点,其中有4个位点可进行PCR-RFLP检测,分别为BstH2I-RFLP,PspEI-RFLP,Eco32I-RFLP和BcnI-RFLP,而在HIF1A和EPAS1扩增片段中未获得可进行PCR-RFLP检测的多态位点。在2个实验猪群中对VEGFA基因的4个多态位点进行基因型及等位基因频率分析,结果表明:VEGFA基因的BstH2I、Eco32I、PspEI 3个多态位点在梅山猪群体中A等位基因占优势,BcnI多态位点在梅山猪群体中B等位基因占优势,而在法系大白猪群体中VEGFA基因的BstH2I、Eco32I、PspEI和BcnI 4个多态位点的相关基因型和等位基因分布极不均衡。BstH2I和PspEI多态位点相关的A等位基因频率在法系大白猪群体中为0,而Eco32I和BcnI多态位点相关的A、B等位基因频率在法系大白猪群体中不到1%。单倍体型推断结果表明VEGFA基因4个多态位点在梅山猪群体中共存在10种单倍体型,而在法系大白猪群体中只存在2种单倍体型BBBA和BBAB,且只有一个个体的单倍体型是BBAB。通过性状关联分析,发现梅山猪群体中VEGFA基因单倍体型BBBA对初产总仔数影响显著(P<0.05);单倍体型BABA对经产总仔数和经产活仔数影响显著(P<0.05);单倍体型BBAA对初生窝重影响显著(P<0.05)。使用辐射杂种克隆板将基因VEGFA定位于SSC7q11-12;基因HIF1A定位于SSC1;基因EPAS1定位于SSC3q11-14。而所获得的基因HIF1A和EPAS1的扩增片段无PCR-RFLP可识别的SNP位点。研究结果表明猪VEGFA基因具有成为影响猪繁殖性状分子遗传标记的潜力。更多还原

【Abstract】 The aim of this work was to try to discover polymorphisms and study the effects of the swine genes VEGFA,HIF1 A and EPAS1 on reproductive traits.After alignment of sequences of different individuals,27 potential SNPs were detected.Four(g.1438C>G,g.1811C>G,g.6973G>T and g.12532C>T)of these SNPs can be detected by PCR-RFLP using digestion of the PCR fragment with BstH2Ⅰ,PspEⅠ,Eco32Ⅰ and BcnⅠ in the intron in the porcine VEGFA gene.The other SNPs cannot be recognized by PCR-RFLP.The alleles of four SNPs were distributed with obvious difference in allele frequency between Meishan pig and French Large White pig.The g.6973G>T polymorphism of the VEGFAgene showed significant correlation with TNB and NBA in later parities(P<0.05)in Meishan pigs.Ten VEGFA haplotypes(GTCT,CGCC,GTGC,CGCT,CGCC,GGGC,GGCT,GGGT,GTCC and CTGT)were detected in Meishan pigs,while only two haplotypes(GTGC and GTCT)in French Large White pigs.The results showed that GGGC haplotype had ignificant effects on TNB/NBA in second and subsequent litters(P<0.05).GTGC haplotype had a significant effect on TNB in first litter(P<0.05)and GTCC had a significant effect on litter born weight(P<0.05).According to IMpRH,VEGFA was assigned to SSC7q11-12,HIF1 A was assigned to SSC1 and EPAS1 was assigned to SSC3q11-14.The results suggest that VEGFAgene associated with swine reproductive traits and further investigation will be necessary to illustrate the underlying mechanism.更多还原