Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro.

摘要: A HindII restriction fragment comprising the Escherichia coli lac regulatory region and genetic information for alpha peptide of beta-galactosidase (beta-D-galactosidegalactohydrolase, EC. 3.2.1.23) has been inserted into 1 10 Bsu I cleavage sites M13 by blunt end ligation. stable hybrid phage was isolated identified its ability to complement function. Further characterization includes retransformation studies, agarose gel electrophoresis, DNA-DNA hybridization, heteroduplex mapping. The insertion point localized at 0.083 map unit on thewild-type circular map-i.e., within intergenic region. results prove that part is nonessential can be used as a cloning vehicle.