反向PCR扩增问号赖型钩体重组质粒pDJH_2插入片段  

Use of Inverse Polymerase Chain Reaction to Amplify DNA Adjacent to the Insert Fragment of pDJH_2 of L.Interrogans Serovar Lai

作　　者：刘洪斌[1] 戴保民[2] 

机构地区：[1]宁波大学医学院,浙江宁波315211 [2]华西医科大学钩体病研究室,四川成都610041

出　　处：《宁波大学学报（理工版）》1999年第3期66-68,共3页Journal of Ningbo University：Natural Science and Engineering Edition

摘　　要：钩端螺旋体赖型赖株（Ｌ．ｉｎｔｅｒｒｏｇａｎｓＳｅｒｏｖａｒｌａｉ）ＤＮＡ分别经ＢａｍＨＩ，ＨｉｎｄⅢ和ＰｓｔⅠ３种内切酶酶切完全消化，通过设计１对引物，反向ＰＣＲ扩增问号赖型钩体重组质粒ｐＤＪＨ２插入片段两端的未知序列．结果显示不同酶切环化连接，经反向ＰＣＲ扩增均可得２条大小相等的扩增带．结果提示反向ＰＣＲ技术可以用于问号赖型钩体重组质粒插入片段两端的未知序列的研究，为获得钩体完整基因序列提供新的途径．DNA of L.interrogans Serovar lai was completely digested by Bam HI,Hind III and Pst I.respectively,then circularized by T4 ligase,and amplified by inverse PCR No matter whatever restrictionenzymes used, the amplification results were the same-two amplificahon bands(450 hp and 250 bP).Theresults indicate that inverse PCR can be used to study the DNA adjacent to a region of knownsequence and to play a role in searching for the complete gene of L.interrogans serovar lai.

关 键 词：钩端螺旋体 反向PCR 重组质粒 pDJH2 钩体病 

分 类 号：R532.12[医药卫生—内科学] S852.63[医药卫生—临床医学]