ISSN:
1617-4623
Keywords:
DNA polymerase III holoenzyme
;
χ subunit
;
holC gene
;
Protein overexpression
;
Codon usage
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract We have identified a previously reported open reading frame (ORF13) that maps between pepA and valS at 96.6 centisomes of the Escherichia coli genome as the structural gene for the χ subunit of DNA polymerase III holoenzyme. This conclusion is supported by a perfect match of the amino-terminal 24 residues of χ with the DNA sequence of ORF13 and a demonstration that ORF13 directs expression of a protein that co-migrates with authentic χ on SDS-polyacrylamide gels. ORF13, designated holC, was isolated from the E. coli chromosome and inserted into a tac promoter-based expression plasmid to direct production of the χ subunit to 5–7% of the total soluble protein. The 3′ end of holC was sequenced to resolve discrepancies between two published versions.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00284693
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