小型猪葡萄糖转运子4外显子4a的基因多态性分析

Polymorphism Analysis of GLUT4 Gene Exon 4a in Three Minipig Strains

目的分析我国特有的3个小型猪品系巴马小型猪、中国农大小型猪、五指山小型猪葡萄糖转运子4基因外显子4a的单核苷酸多态性(SNPs)分布特点,为我国小型猪在糖尿病和代谢性疾病研究中提供基础资料。方法以3个品系小型猪基因组DNA为模板,应用特异性引物进行PCR扩增,扩增产物纯化测序,进行BLAST比对分析。结果在小型猪GLUT-4基因外显子4a上有2个SNP位点:SNP1:GCT→GCC(Ala133 Ala),3个品系均发生了变化,均为纯合突变,其突变率为(22/22,100%)。SNP2:GGC→GGT(Gly146 Gly),巴马小型猪突变率为(6/6,100%),均为纯合突变;五指山小型猪突变率为(6/6,100%),均为纯合突变;中国农大小型猪突变率为(10/10,100%),其中包括6例纯合突变(6/10,60%)和4例杂合突变(4/10,40%)。结论SNPs1,在所有测定的小型猪品系中检出,这可能是所测小型猪的共有特征。SNPs2可能是小型猪品系之间的差异。

Objective To analyze the single nucleotide polymorphism （SNPs） of glucose transporter-4 gene exon4a in three Chinese minipig strains： Bama minipig, Wuzhishan minipig and Nongda minipig, to provide basic data for their application in researches on diabetes mellitus and metabolic diseases. Methods Genomic DNA was extracted from blood samples of the minipigs, and served as templates for target gene segment amplification and selection by polymerase chain reaction （PCR）. The PCR products were sequenced and analyzed for documenting single-nucleotide polymorphisms （SNPs）. Results Two SNPs were found in the target region. One was GCT→GCC （Ala133 Ala）, which was discovered from all three minipig strains and all were homozygous mutations. The other was GGC→GGT （Gly146 Gly）, which was found from Bama minipig strain with all homozygous mutations, Wuzhishan minipigs with all homozygous mutations, and Nongda minipig including heterozygous and homozygous mutations. Conclusion SNP1 are detected from all the three miniature pigs. It may be a specific mutation in Chinese native minipigs. It may be a common trait among the three minipig strains. SNP2 may be a difference among minipig strains.