【摘要】 EcoRI酶解的AsGV-XJ DNA和质粒pUB 110 DNA,经T4 DNA连接酶连接后,转化枯草芽孢杆菌BR 151感受态细胞,涂布在加有新霉素（5μg/ml）的完全培养基上。用琼脂糖凝胶电泳快速法检测抗新霉素转化子中的重组质粒。在388个转化子中得到12个较pUB110 DNA分子量大的重组质粒。所有重组质粒均可与³²PdCTP标记的EcoRI酶解AsGV-XJDNA片段的探针进行分子杂交。通过琼脂糖凝胶电泳对重组质粒中插入AsGV-XJ DNA片段进行了测定。更多还原

【Abstract】 Fragments of the DNA of Agrotissegetum granulosis virus isolated fromXinjiang, China (designed as AsGV-XJ)can be cloned in Bacillus subtilis. TheAsGV-XJ DNA and pUB 110 DNA weredigested with EcoRI restriction endonuclea-se, ligated with T4 DNA ligase, then tran-sformed into B. subtilis BR 151 competentcells. The transformants were plated ontoBPY medium containing neomycin 5μg/ml.The molecular weights of the 12 recom-binant plasmids isolated from 388 transfor-mants were larger than that of pUB 110plasmid determined by rapid electrophore-sis in slab agarose gels. All of recombinantDNA clones were identified as the frag-ments of AsGV-XJ DNA digested withEcoRI by the Southern dot and blot tran-sfer hybridization. Size of the fragmentsinserted into recombinant plasmids wereranged from 0.2 to 4 kb.更多还原