Mutation Surveyor在插入/缺失杂合子突变分析中的应用  

Application of Mutation Surveyor in analyzing heterozygous InDel

作　　者：宋雪梅[1] 姜俊芳[1] 蒋永清[1] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,浙江杭州310021

出　　处：《浙江农业学报》2012年第6期961-968,共8页Acta Agriculturae Zhejiangensis

基　　金：浙江省重大科技专项(2010C12007);浙江省自然科学基金(Y308497);国家科技支撑项目(2008BADB2B04)

摘　　要：结合实例简述了Mutation Surveyor软件在碱基替换和插入/缺失突变,尤其是插入/缺失杂合子突变研究中的应用。以浙江省余杭湖羊场和屠宰场144头湖羊为试验材料,采用PCR-SSCP和测序的方法检测干扰素刺激基因(ISG15)基因5'侧翼区序列,并利用Mutation Surveyor软件分析其遗传多态性。PCR-SSCP检测结果显示ISG15基因5'侧翼区PCR-SSCP带型非常复杂,无法直接判定个体的基因型。利用Mutation Survey-or软件分析发现湖羊ISG15基因5'侧翼区存在152 C/T,158 dupC和239 C/T 3种突变。其中152 C/T C等位基因频率为0.83,为低度多态的分子标记;158 dupC等位基因频率为0.5125,239 C/T C等位基因频率为0.7675,均为中度多态的分子标记。除239 C/T外,152 C/T和158 dupC均处于Hardy-Weinberg平衡状态(P>0.05)。试验结果显示Mutation Surveyor软件在分析插入/缺失杂合子突变的测序结果时有较大优势,可用于碱基替换和插入/缺失SNPs的通量筛选和分析。The present study aimed to show the application of Mutation Surveyor in analyzing nucleotide substitutions and InDel with an instance. The polymorphisms in ISG15 5′ flanking sequences were analyzed for 144 Hu sheep from Yuhang Hu sheep farm and slaughterhouse by polymerase chain reaction-single strand confornmtion polymorphism （PCR-SSCP） technique, DNA sequencing and Mutation Surveyor software. PCR-SSCP analysis showed 1SG15 5′ flanking sequences bands were too complex to determine genotype directly. The analysis result of Mutation Surveyor found there were three polymorphisms, 152 C/T, 158 dupC and 239 C/T, in ISG15 5 ′flanking region. The frequen- cy of allele C in loci 152 was 0. 83 showing low polymorphism （PIC = 0. 2424）. The frequency of allele dupC in lo- ci 158 was 0. 5125 showing mediate polymorphism （ PIC = 0. 3748 ）. The frequency of allele C in loci 239 was O. 7675 showing mediate polymorphism （PIC = 0. 2937.）. Except 239 C/T, the gene frequencies and genotype fre- quencies of Hu sheep 152 C/T and 158 dupC tested have reached the Hardy-Weinberg equilibrium （P 〉 0. 05）. Mu- tation Surveyor software has greater advantage in analysis of nucleotide substitutions, especially InDel sequencing da- ta. So Mutation Surveyor software can be used for SNPs high-throughput screening and analysis.

关 键 词：MUTATION SURVEYOR INDEL ISG15 湖羊 

分 类 号：S823.2[农业科学—畜牧学]