Abstract:

Mutation scanning of Strongylocentrotus intermedius genes was conducted by high-resolution melting curve (HRM) analysis technology. 26 alternative SNP loci were screened out. Among them, 16 were base substitution accounting for 61.54%, 6 were base deletion or insertion accounting for 23.08%. The number of missing bases was ranging from 1 to 4. 4 are fuzzy base substitution type and tri-state loci accounting for 15.38%. According to these figures, SNP frequency distribution in this gene is about 2.85%, and 18 loci in the coding region, among which 2 are missense mutations. We analyzed 5 SNP loci using small amplicon genotyping in 60 individuals from 2 populations from different regions. The results indicated that all 5 loci had 2 alleles, the minor allele frequency ranged from 0.150 0 to 0.500 0, and SILyz-7, SILyz-15 and SILyz-20 had large differences in minor allele frequency between 2 populations. It could be speculated that HRM is an efficient and reliable SNP analysis method, and the BNP marker could be used for studying Strongylocentrotus intermedius genetics and breeding.

Key words: Strongylocentrotus intermedius, lysozyme, high resolution melting (HRM), single nucleotide polymorphism (SNP)

摘要：

采用高分辨率熔解曲线分析技术（HRM）对虾夷马粪海胆溶菌酶基因进行突变扫描，筛选出26个SNP候选位点。其中，碱基置换类型位点16个，占位点总数的61.54%；碱基插入或缺失的位点6个，缺失的碱基数为1~4个不等，占位点总数的23.08%；碱基置换类型模糊以及三态位点4个，占位点总数的1538%。SNP在虾夷马粪海胆溶菌酶基因的cDNA中的分布频率约为2.85%，位于编码区的位点有18个，其中2个为错义突变。应用小扩增子基因分型法对5个SNP位点在2个不同地域群体的60个样本中进行分型分析，结果显示：5个SNP位点均含有2个等位基因，次等位基因频率分布范围为0.150 0~0.500 0，位点SILyz-7、SILyz-15和SILyz-20在两个群体间的次等位基因频率相差较大。研究结果表明，HRM是一种高效便捷的SNP分析方法，所筛选出的SNP标记可用于虾夷马粪海胆遗传育种的研究。

关键词: 虾夷马粪海胆;溶菌酶;高分辨率熔解曲线分析;SNP