含有来自噬菌体T4的DNA片段的噬菌体λ和质粒DNA分子的体外构建。

In vitro construction of bacteriophage lambda and plasmid DNA molecules containing DNA fragments from bacteriophage T4.

作者信息

Velten J, Fukada K, Abelson J

出版信息

Gene. 1976;1(1):93-106. doi: 10.1016/0378-1119(76)90009-3.

Abstract

Restriction endonucleases EcoRI and HindIII generated fragments of T4 cytosine-containing DNA were inserted into bacteriophage vector lambdagtSuIII and plasmid vectors pMB9 and pBR313. Resulting clones were screened for hybridization with 32P labeled T4 tRNA. Recombinant bacteriophages and plasmids were isolated which contained a T4 fragment coding for T4 RNA species 1 and 2 and T4 tRNA Arg. Selected lambda-T4 hybrid bacteriophages were grown to high titer and their DNA analyzed by gel electrophoresis.

摘要

用限制性内切酶EcoRI和HindIII切割含胞嘧啶的T4 DNA，产生的片段被插入到噬菌体载体λgtSuIII以及质粒载体pMB9和pBR313中。用32P标记的T4 tRNA筛选所得的克隆。分离出含有编码T4 RNA种类1和2以及T4 tRNA Arg的T4片段的重组噬菌体和质粒。挑选出的λ-T4杂交噬菌体增殖至高滴度，并通过凝胶电泳分析其DNA。