Cloning and identification of the product of the dnaE gene of Escherichia coli

Publisher Website
Google Scholar
Abstract
The dnaE gene of E. coli was successively cloned into pBR322, resulting in a plasmid that contains 4.6 kilobases of E. coli DNA. This plasmid can complement a dnaE temperature-sensitive mutation. A restriction map of the dnaE gene and the surrounding 10.7-kilobase region of the E. coli chromosome was determined. A unique HindIII restriction endonuclease site within the cloned segment of DNA was identified as a site required for expression of the dnaE gene. By using the maxicell plasmid-directed protein synthesizing system, it was demonstrated that dnaE codes for the .alpha. subunit of DNA polymerase III.