猪POU1F1基因部分序列变异和同源性分析

Variation and Homologous Analysis of Parts of Pig POU1F1 Gene

对长白、杜洛克、约克夏、姜曲海、梅山和香猪等六个猪种的POU1F1基因第四、第五和第六外显子分别进行PCR扩增 ,并对含有第四、第六外显子的PCR产物和含有第五外显子的克隆产物进行测序。结果表明 :六个猪种中 ,POU1F1基因的第四外显子存在碱基突变 ,为T→C。对该序列进行NlaⅢ酶消化 ,产生两种不同的基因型(GG和HH) ;而第五和第六外显子则高度保守 ,未发现任何突变。将人POU1F1基因第四外显子、POU同源区核苷酸编码序列和氨基酸序列 ,分别与猪、小鼠、牛的POU1F1基因相应的核苷酸序列和氨基酸序列进行同源性比较 ,结果发现 :人与猪、小鼠、牛的POU1F1基因第四外显子的核苷酸同源性分别高达 93 9%、86 7%、92 1% ,而由第四外显子编码的部分POU特异区的氨基酸序列则完全一致 ;人与猪、小鼠、牛POU同源区的核苷酸同源性分别为91 4 %、85 1%、87 9% ,氨基酸同源性分别为 96 6 %、94 8%、90 2 %。这说明在哺乳动物中 ,其POU1F1蛋白的POU同源区和由第四外显子编码的POU特异区部分是高度保守的 ;猪可作为实验动物 ,建立人类相关疾病模型 。

By PCR approach,exon4,exon5 and exon6 of POU1F1 gene of Landrace,Duroc,Yorkshire,Jiangquhai、 Meishan,and Xiangzhu are amplified,and PCR products contained exon 4,exon6 and the cloning product of exon5 were sequenced. The results showed that exon4 of POU1F1 gene had the Nla Ⅲ polymorphisms characteristics,the mutation was T→C.The sequences were digested by Nla Ⅲ,and two different genotypes(GG and HH) were produced; but exon 5 and exon 6 of POU1F1 gene were high conservation in the six pig breeds. The result of homologous analysis showed that the homologous rate of nucleotide sequences of the exon4 in human,pig,mouse and cow was 93.9%、86.7%、92.1% respectively and the amino acid sequence of POU specific domain encoded by exon4 of POU1F1 gene in human,pig,mouse and cow was identical. And the result of homologous analysis of nucleotide sequences encoding POU1F1 POU-homeo domain in human,pig,mouse,cow was 91.4%、85.1%、87.9% respectively,the homologous rate of the amino acid sequence of POU-homeo domain was 96.6%、94.8%、90.2% respectively. This showed that the sequences of the POU1F1 POU-homeo domain and the POU-specific domain encoded by exon4 in mammals were high conservation,pigs could be used to establish animal models of related diseases,providing scientific foundation and serving for human medicine.