摘要
本试验采用了PCR-SSCP和PCR-RFLP技术,对不同猪种(长白、杜洛克、约克夏、姜曲海、梅山和香猪等)的POU1F1基因第四、第五和第六外显子的序列变异和第四外显子、第三内含子的序列多态性进行研究。同时分析了第三内含子的不同基因型与杂交商品猪生长性能之间的关系。试验结果表明:
1.在不同猪种中,POU1F1基因的第五和第六外显子没有出现任何多态性;POU1F1基因第四外显子在Nla Ⅲ酶切位点存在多态性,由此产生两个等位基因(G,H)。国外优良的瘦肉型猪种的等位基因H的频率较高,中国地方猪种的等位基因H的频率较低。人、猪、小鼠和牛的POU1F1蛋白的POU同源区和由第四外显子编码的POU特异区高度保守。
2.猪POU1F1基因第三内含子在Msp Ⅰ酶切位点上表现出了丰富的多态性,而且不同猪种在这个位点上的基因频率差异显著。国外瘦肉型猪种的等位基因D的频率较高,而国内高繁殖力和脂肪型猪种的等位基因D的频率较低。
3.在分析杂交商品猪POU1F1基因第三内含子的多态性和生长性能的相关性时发现,DD基因型个体在不同日龄段的体重(除初生重外)和日增重分别与其它基因型个体相同日龄段的体重和日增重之间差异显著(P<0.05)或极显著(P<0.01)。
4.分别将不同物种POU1F1蛋白的POU区域的氨基酸序列及其核苷酸编码序列进行同源性比较分析,人与猪POU1F1蛋白的POU区域的同源性显著高于人和其它动物POU1F1蛋白的POU区域的同源性,提示可用猪作为实验动物,建立相关的人类动物疾病模型,为医学研究提供参考依据。
This research was conducted to analyse the sequence variations in partial region of pig POU1F1 gene, including exon4, exon5, exon6 and intron3, by using PCR-RFLP, PCR-SSCP and sequencing techneque. Six pig breeds (Landrace, Duroc, Yorkshire, Jiangquhai, Meishan and Xiang) were selected as experimental animal. Also a commercial pig population growth performance was correlated with the polymorphism of intron3. The results were shown as follows:
1. No polymorphism was found in exonS and exon6 of POU1F1 gene in all breeds. A point mutation but amino acid substitute was identified in exon4. Allele (G, H) frequencies in this restriction site in different pig breeds distributed significantly, with high H allele frequency in foreign meat pigs and low in Chinese local breeds. While among human, pig, mouse and cow, POUHD and POUso encoded by exon4 were high conservation.
2. A Msp I polymorphism reported previously was identified in intron3. There was a significant difference of allele frequencies in this site among different breeds as well. The D allele frequency was high in foreign meat pigs, while low in Chinese local fat pigs.
3. According to analysis of correlation between growth performance and genotypes in commercial pigs, body weight (expect birth weight) and daily gain in different stages between DD genotype and other genotypes differed significantly (P<0.05) or extremely significantly (P<0.01), respectively.
4. By comparing homologous analysis of nucleotide sequences encoding POU domain of POU1F1 protein and amino acid sequences in different species respectively, the
homologous rate between human and porcine POU domain of POU1F1 protein was significantly higher than that of human and other animals POU domain of POU1F1 protein, so pigs can be used to establish animal models of related human diseases, providing scientific foundation and serving for human medicine.
引文
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