虾夷马粪海胆溶菌酶基因SNP标记的开发及多态性分析  

Development and Polymorphism Analysis of SNP Marker in LYZ of Strongylocentrotus intermedius

作　　者：丛晓霏[1] 丁君[1] 常亚青[1] 

机构地区：[1]大连海洋大学,农业部北方海水增养殖重点实验室,辽宁大连116023

出　　处：《中国农业科技导报》2012年第5期136-144,共9页Journal of Agricultural Science and Technology

基　　金：辽宁省“百千万人才工程”资助项目(2008921059);辽宁省科技计划(2007203004);国家自然科学基金(30972269);国家863计划项目(2012AA10A412);辽宁省渔业厅项目(200916)资助

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。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.

关 键 词：虾夷马粪海胆 溶菌酶 高分辨率熔解曲线分析 SNP 

分 类 号：Q959.268.3[生物学—动物学]