本试验旨在研究内皮型一氧化氮合酶(eNOS)基因exon8多态性与中国美利奴羊布鲁氏菌病易感性的相关性。利用生物信息学方法对人和中国美利奴羊eNOS基因exon8序列进行比对,并对NCBI SNP数据库上人eNOS基因exon8多态性进行了统计分析。通过PCR-SSCP对101只中国美利奴羊阴性样本和61只中国美利奴羊阳性样本eNOS基因exon8的多态性进行检测,然后对不同等位基因进行PCR产物测序,旨在确定该基因exon8多态性位点,并对SNP位点的等位基因频率、基因型频率进行统计分析。在eNOS基因exon8序列142bp处检测到一个新的SNP位点(ss974768653:A142G),该位点在病例组和对照组之间的等位基因频率及各基因型间不存在显著差异性(P〉0.05)。eNOS基因exon8A142G多态性位点与中国美利奴羊布鲁氏菌病易感性可能无相关性。
The association of Chinese Merino sheep eNOSgene exon8 polymorphisms with brucellosis susceptibility was studied in this research.The sequences of human and Chinese Merino sheep eNOSgene exon8 were aligned by the bioinformatics methods,and the polymorphisms of human eNOSgene in the NCBI SNP database were statistically analyzed.The polymorphisms of101 Chinese Merino sheep negative samples and 61 Chinese Merino sheep positive samples were detected by PCR-SSCP method,and then the PCR products of different alleles were sequenced,aiming to determine exon8's polymorphism loci,and the allele frequency and genotype frequency of SNP loci were statistically analyzed.A novel SNP locus(ss974768653:A142G)was detected at the 142 bp of eNOSgene exon8,and the locus had no difference in allele frequency and each genotype between negative and positive groups(P〉0.05).There might be no correlation between the A142 Gpolymorphism locus of Chinese Merino sheep eNOSgene exon8 and brucellosis susceptibility.