Wu Fen, Zhang Wei, Song Qian-Qian, Li Hai-Hong, Xu Ming-Shu, Liu Guo-Liang, Zhang Jin-Zhi
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Zhejiang Qinglian Food Company Limited, Jiaxing 314000, China.
Asian-Australas J Anim Sci. 2019 Aug;32(8):1104-1111. doi: 10.5713/ajas.18.0827. Epub 2019 Jan 4.
The aim of this study was to detect single nucleotide polymorphisms (SNP) of G protein-coupled receptor 54 (GPR54) gene and explore association of this candidate gene with reproductive traits in Jiaxing Black sows.
Six pairs of primers of the gene were designed to amplify all exons thus sequences of which were detected by means of direct sequencing and then SNP loci were scanned. The effects of SNPs on total number of piglets born (TNB), number of piglets born alive (NBA), number of still born piglets (NSB), and litter weight at birth (LWB) of Jiaxing Black sows were analyzed.
Three SNP loci, including T3739C, C3878T and T6789C, were identified via comparison of sequencing and two genotypes (AB, BB) at each SNP site were observed. T3739C resulted in the change of amino acid (Leu→Pro) in corresponding protein, and C3878T resulted in synonymous mutation (Ile→Ile). Statistical results demonstrated that allele B was the preponderant allele at the three SNP loci and Genotype BB was the preponderant genotype. Meanwhile, Chi-Square test of these three SNPs indicated that all mutation sites fitted in Hardy-Weinberg equilibrium (p>0.05). For GPR54-T3739C locus, Jiaxing Black sows with genotype BB had 1.23 TNB and 1.28 NBA (p<0.01) that were more than those with genotype AB, respectively. Jiaxing Black sows that had the first two parities with genotype BB had additional 2.23 TNB, 2.27 NBA (p<0.01), and 1.94 LWB (p<0.05) compared to those with genotype AB, respectively. However, for other two loci, no significant difference was found between TNB, NBA, NSB, and LWB, and different genotypes of Jiaxing Black sows.
In conclusion, the polymorphisms of GPR54-T3739C locus were significantly associated to TNB, NBA, and LWB and could be used as a potential genetic marker to improve reproductive function of Jiaxing black sows.
本研究旨在检测G蛋白偶联受体54(GPR54)基因的单核苷酸多态性(SNP),并探讨该候选基因与嘉兴黑母猪繁殖性状的关联。
设计该基因的6对引物,扩增所有外显子,通过直接测序检测其序列,进而扫描SNP位点。分析SNP对嘉兴黑母猪产仔总数(TNB)、产活仔数(NBA)、死胎数(NSB)和初生窝重(LWB)的影响。
通过测序比对鉴定出3个SNP位点,分别为T3739C、C3878T和T6789C,每个SNP位点均观察到两种基因型(AB、BB)。T3739C导致相应蛋白质中氨基酸发生变化(Leu→Pro),C3878T导致同义突变(Ile→Ile)。统计结果表明,在这3个SNP位点上,等位基因B为优势等位基因,基因型BB为优势基因型。同时,对这3个SNP进行卡方检验表明,所有突变位点均符合哈迪-温伯格平衡(p>0.05)。对于GPR54-T3739C位点,基因型BB的嘉兴黑母猪TNB为1.23头,NBA为1.28头(p<0.01),分别高于基因型AB的母猪。前两胎基因型为BB的嘉兴黑母猪与基因型AB的母猪相比,TNB额外增加2.23头,NBA额外增加2.27头(p<0.01),LWB额外增加1.94千克(p<0.05)。然而,对于其他两个位点,嘉兴黑母猪不同基因型在TNB、NBA、NSB和LWB方面未发现显著差异。
综上所述,GPR54-T3739C位点的多态性与TNB、NBA和LWB显著相关,可作为提高嘉兴黑母猪繁殖性能的潜在遗传标记。