Cloning, Sequencing, and Enhanced Expression of the Dihydropteroate Synthase Gene of Escherichia Coli MC4100

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1992 Sep 1

PMID 1522070

Citations 40

Authors

W S Dallas

J E Gowen

P H Ray

M J Cox

I K Dev

Affiliations

Soon will be listed here.

Abstract

The Escherichia coli gene coding for dihydropteroate synthase (DHPS) has been cloned and sequenced. The protein has 282 amino acids and a compositional molecular mass of 30,314 daltons. Increased expression of the enzyme was realized by using a T7 expression system. The enzyme was purified and crystallized. A temperature-sensitive mutant was isolated and found to express a DHPS with a lower specific activity and lower affinities for para-aminobenzoic acid and sulfathiazole. The allele had a point mutation that changed a phenylalanine codon to a leucine codon, and the mutation was in a codon that is conserved among published DHPS sequences.

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