【作者】 李兴安； 牛庆生； 薛运波； 陈东海； 张发；

【Author】 LI Xing-an;Niu Qing-sheng;XUE Yun-bo;CHEN Dong-hai;Zhang Fa;Jilin Provincial Institute of Apicultural Science;

【机构】 吉林省养蜂科学研究所；

【摘要】 蜜蜂基因组微卫星DNA或简单序列重复DNA (simple sequence repeats,SSR或SSRs)基因座,是绘制西方蜜蜂基因组遗传图谱的主要界标,是组装西方蜜蜂基因组序列图谱的架构。分子标记应用于蜜蜂身份鉴定是蜜蜂育种研究的基础工作之一。为了揭示蜜蜂基因组SSR基因座的可利用性,本文通过微流体DNA芯片,在基因组水平对欧洲黑蜂雄蜂基因组连锁群-I的273个SSR基因座PCR产物进行了全扫描。结果显示,在每一个SSR基因座PCR产物的微流体胶分离谱图中,SSR侧翼序列引物自身扩增产物、SSR扩增产物和非SSR扩增产物被表征为有差异的优势扩增信号测定值和非优势扩增信号测定值;所有SSR基因座PCR产物的微流体胶分离谱图呈现5种类型,这包括SSR侧翼序列引物优势扩增类型(第I种),SSR优势扩增类型(第Ⅱ种),SSR侧翼序列引物和SSR均优势扩增类型(第Ⅲ种),SSR和非SSR均优势扩增类型(第Ⅳ种),以及SSR侧翼序列引物、SSR和非SSR均优势扩增类型(第V种)。结果进一步显示,在SSR优势扩增类型中,14个SSR基因座PCR产物明显地符合3个测定值限量值(或阈值),即PCR优势扩增产物质量≥10ng/uL、PCR优势扩增产物质量与PCR扩增总产物质量之比≥0.7以及平均非PCR优势扩增产物数量≤3。这提示,按照PCR扩增产物的微流体胶分离谱图特征,西方蜜蜂基因组连锁群-I的SSR基因座可以划分为PCR特异性扩增SSR位点和PCR非特异性扩增SSR位点;西方黑蜂基因组连锁群-I的14个SSR基因座可能成为西方蜜蜂身份鉴定的可利用SSR分子标记。更多还原

【Abstract】 Microsatellite or simple sequence repeat(SSR or SSRs) loci in genome were served as key mapping sites in genetic map.The genotyping map provides a framework of SSR site to set up genomic assembly.Molecular markers in the field of bee breeding research have been developed into DNA barcodes to be informative for identification of honeybee species.To reveal availability of genomic SSR in future,we carried out an assay of 273 SSR loci in linkage group-Ⅰ of honeybee(Apis mellifera mellifera) drones at genome level using microfluidic DNA chip in high through-put screen.It is shown from patterns of both SSR electrophorogram and electro-chromatogram in microfluid matrix that PCR amplicons of any SSR locus from DNA samples was indicated as measurement values of dominant and dispersal signals,including SSR flank primers,specific SSRs and nonspecific SSRs.Furthermore,it is shown from the patterns that PCR amplicons of all SSR loci from DNA samples were presented as five types,including dominant SSR flank primers(Ⅰ),dominant SSRs(Ⅱ),dominant SSR flank primers and SSR(Ⅲ),dominant SSR and nonspecific SSRs(Ⅳ),and dominant SSR flank primers,SSR and nonspecific SSRs(V).When thresholds for the measurement value are set up simultaneously as dominant PCR amplicons≥ 2 ug/uL,ratio of dominant PCR amplicons to total PCR amplicons≥0.7,and the number of dispersal PCR amplicons≤3,14 available SSR loci are differentiated significantly from the 273 SSR loci.In suggested from the study that 273 SSR loci in linkage group-Ⅰ of Apis mellifera could be classified as two category of SSR sites,specific and nonspecific PCR amplification,and 14 SSR sites of specific PCR amplification could be developed into DNA barcodes for identification of Apis mellifera.更多还原