AFLP法构建布鲁氏菌基因组DNA指纹图谱  

Construction of genomic DNA fingerprinting of Brucella by AFLP

作　　者：骆利敏[1] 李明[1] 芮勇宇[2] 陈瑶[1] 王文敬[1] 

机构地区：[1]南方医科大学生物技术学院,广州510515 [2]南方医科大学附属南方医院检验科

出　　处：《中国人兽共患病学报》2006年第1期15-17,共3页Chinese Journal of Zoonoses

基　　金：973课题2002CB513206基金资助

摘　　要：目的采用扩增片段长度多态性(AFLP)分子遗传标志技术,分析布鲁氏菌基因组DNA多态性。方法提取布鲁氏菌基因组DNA,经EcoRI/MseI酶切并与相应的人工接头连接后,使用选择性引物进行PCR扩增。结果经变性聚丙烯酰胺凝胶电泳检测,成功构建出多态性丰富、重复性好的布鲁氏菌DNA指纹图谱。结论AFLP法有望成为一种独立的切实可行的方案,在布鲁氏菌的多态性研究和流行病学调查中发挥作用。To analyse the genetic polymorphism of Brucella by the molecular genetic marker technique with amplified fragment length polymorphism （AFLP） , genetic DNA extracted from Brucella was digested by EcoRI in combination with MseI and ligated with EcoR I and Mse I adapters respectively. Meanwhile, the polymerase chain reactions were performed with E-ACA and MCAG selective primer pairs. Then the distinct, reproducible genomic DNA fingerprints were constructed using denatured polyacrylamide gel electrophoresis assay. Eighteen Brucella strains were divided into eight groups and could be differentiated on the level of species. The results suggested that AFLP could be a powerful tool in polymorphism study and molecular epidemiology investigation of Brucella .

关 键 词：布鲁氏菌 扩增片段长度多态性 DNA捂纹图谱 

分 类 号：R378[医药卫生—病原生物学]