从江香猪MC4R基因多态性及生物信息学分析

Polymorphic detection and bioin for matics analysis of MC4R gene in Congjiang Xiang pig

[目的]旨在对3个群体猪MC4R基因进行SNPs筛选,为从江香猪选种选育提供一定的理论基础。[方法]以从江香猪作为研究对象,野猪×从江香猪二元杂交猪和杜×大×长外三元杂交猪作为对照,构建品种DNA池,PCR扩增MC4R基因外显子、内含子和3’非编码区序列,采用直接测序法对3个群体的MC4R基因进行单核苷酸多态性检测,利用生物信息学软件预测不同多态性位点对MC4R基因mRNA二级结构和蛋白质二级结构的影响。[结果]在3个群体中共检测出10个SNPs位点,其中,C860T、G1392A和G1577A位于第2外显子区;G1699A、T1702A、A1870C、G1875A、C2025T、G2073A和A2111T位于3’非编码区序列。C860T为同义突变,G1392A和G1577A为错义突变,分别导致了精氨酸变为组氨酸、天冬氨酸变为天冬酰胺。经分析软件预测,处于第二外显子中的3个SNPs位点对MC4R基因的mRNA二级结构、蛋白质二级结构有一定的影响。[结论]DNA池结合测序技术检测到MC4R基因外显子、内含子和3’非编码区序列共10个SNPs。

[Objective]This experiment was per for med to screen the single nucleotide polymorphisms（SNPs） in the three groups of pigs＇ MC4R gene which could lay the experiment foundation for heredity selection of Congjiang Xiang pigs.[Methods]Congjiang Xiang pigs were selected for research subject,the commercial crossbred pigs and the wild boar × Xiang pigs were contrast animals respectively.constructingDNA pools amplified all sequences of all extrons and introns and 3＇noncoding regions.using the technology of direct sequencing detected the single nucletide polymorphisms on MC4R gene in three breeds.Through bioin for matics analysis we found that the polymorphisms result in the changes of mRNA secondary structure and protein secondary structure.[Results]The study have totally found10 SNPs in the three group.Among these SNPs,C860 T,G1392A and G1577 A were located in extron2,G1699 A,T1702A,A1870 C,G1875A,C2025 T,G2073A and A2111 T were located in 3＇noncoding regions.the C860 T loci belongs to the same sense mutation while the G1392 A and G1577 loci belong to the missense mutation.The 3 SNPs in extron2 of MC4R gene exert profound impact on the secondary structure of mRNA and protein.[Conlusion]The ten SNPs were found in all extrons and introns and 3＇noncoding regions of MC4R gene by using this technology ofDNA pooling.