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.
出处
《中国农业科技导报》
CAS
CSCD
北大核心
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
关键词
虾夷马粪海胆
溶菌酶
高分辨率熔解曲线分析
SNP
Strongylocentrotus intermedius
lysozyme
high resolution melting （HRM）
single nucleotidepolymorphism （SNP）
