Cloning of pac gene and its flanking regions using mini-Mu bacteriophage.

摘要

An in vitro method for cloning and mapping Escherichia coli genes by means of mini-Mu phage and its application to the penicillin G acylase (pac) gene of E. coli PAC2 strain with its flanking regions is described. The gene was marked by insertion of a fragment bearing kanamycin resistance (Km(r)). Most of Km(r) clones obtained from mini-Mu transductants contained the whole pac gene with its flanking regions. Localization of pac gene to 98.5 min of E. coli PAC2 chromosome was confirmed by an in vivo P1 phage transduction.