“三明治”杂交法对斜纹夜蛾核型多角体病毒DNA克隆株的鉴定

Identification of PINPV--DNA Recombinant Colonies with Sandwich Hybridization

用限制性内切酶HindⅢ将斜纹夜蛾核型多角体病毒DNA切割为A、B、C、D_1、D_2、E、F七个片段,与载体pUC18体外连接后转化大肠杆菌TG_1。通过“三明治“杂交法鉴定,分别获得七个片段的克隆株。但A片段克隆株中外源DNA片段发生了缺失。

The genome of the bacularvirus PINPV was digested completely into sevenfragment (A、B、C、D_1、D_2、E、F) with Hind Ⅲ. These fragments were ligated in vitro with vector pUC18 seperately and used to transform host strainE. coli TG_1. Recombinant colonies were identified with sandwich hybridization.Seven kinds of recombinant colonies containing the PINPV-DNA Hind Ⅲ fra-gments were obtained, but unfortunately the A fragment was less than theoriginal one.