东北黑熊不同基因组DNA模板来源的比较研究  

Study on different sources of U.t.ussuricus genomic DNA template

作　　者：高建伟[1] 鲁承[1] 崔正云[1] 崔成都[1] 贾文影 李刚 

机构地区：[1]延边大学农学院动物医学系,吉林延吉133002 [2]科菲特饲料(齐齐哈尔)有限公司,黑龙江齐齐哈尔161200 [3]长春大政药业科技有限公司,长春130102

出　　处：《东北农业大学学报》2012年第9期90-96,共7页Journal of Northeast Agricultural University

基　　金：校企合作研究项目(延大科合字(2009第34号))

摘　　要：文章主要探讨使用黑熊毛发作为试验材料替代其他生物样品进行遗传多样性研究的可行性,筛选满足试验需求的最佳使用量。以延边白头山制药有限公司养熊场的4头成年东北黑熊为试验材料,分别5次拾取2头黑熊铁笼内散落的带有毛囊的50、100根毛发,对麻醉的2头黑熊抽取全血3 mL以及皮下结缔组织1 g,并分别采用不同方法提取基因组DNA。以微卫星位点ABB12和UT35为引物,设置20、50、100 ng·μL-13种模板浓度进行PCR扩增。结果显示,采用50 ng·μL-1的模板浓度均可获得较好的PCR产物,采用聚丙烯酰胺凝胶电泳对扩增基因具有较好的分辨效果。毛发DNA、血液DNA和组织DNA的PCR产物数量和质量并无明显差异。毛发数量与所提取的DNA浓度有一定关系。利用15个毛发DNA的PCR产物进行聚丙烯酰胺凝胶电泳,均可观察到清晰的条带。研究结果进一步证明毛发DNA可作为东北黑熊遗传多样性研究的试验材料。The research aimed to discuss the feasibility of make the genegic diversity research by using U.t.ussuricus hair wholly as tested materials.With four adult U.t.ussuricus of Yanbian Baitou mountain pharmacectic company,co,ltd bear breeding factory as tested materials,50 and 100 hairs were picked up five times in two bear cages and 3 mL whole blood were taken out and 1g subcutaneous connective tissue were cut to make DNA extraction in different methods.The blood and subcutaneous connective tissue were obtained,when the two bears were narcotized.With microsatellite primers ABB12 and UT35 as primers,3template concentration including 20,50,100 ng.μL-1 were set up to make PCR amplification.Using 50 ng.μL-1 as template concentration could obtain better PCR products,which had better resolving effects on the amplification genes in polyacrylamide gel electrophoresis.The quantity and quality of PCR products among hair,blood and tissue DNA samples had no obvious difference.The hair number had suitable correlation with the concentration of DNA.When the PCR products of 15 whole-hair samples were used in the native polyacrylamide gel electrophoresis,clear bands could be observed.The research results further proved that DNA from whole-hair samples could be taken as the materials for U.t.ussuricus genetic diversity research.

关 键 词：不同基因组DNA模板 微卫星 PCR扩增 

分 类 号：Q756[生物学—分子生物学]