Molecular cloning and characterization of the aroD gene encoding 3-dehydroquinase from Salmonella typhi

S. Servos, S. Chatfield, D. Hone, M. Levine, G. Dimitriadis, D. Pickard, G. Dougan, N. Fairweather, I. Charles

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Abstract

The aroD gene from Salmonella typhi, encoding 5-dehydroquinate hydrolase (3-dehydroquinase), has been cloned into Escherichia coli and the DNA sequence determined. The aroD gene was isolated from a cosmid gene bank by complementation of an S. typhimurium aroD mutant. Analysis of the DNA sequence revealed the presence of an open reading frame capable of encoding a protein of 252 amino acids with a calculated M(r) of 27,706. Comparison of the deduced S. typhi 3-dehydroquinase protein sequence with that elucidated for E. coli revealed 69% homology. Alignment of the S. typhi sequence and equivalent Aspergillus nidulans and Saccharomyces cerevisiae sequences showed that homology was lower, at 24%, but still significant. Use of a minicell expression system demonstrated that a polyclonal antibody raised against E. coli 3-dehydroquinase cross-reacted with its S. typhi counterpart.

Original languageEnglish
Pages (from-to)147-152
Number of pages6
JournalJournal of General Microbiology
Volume137
Issue number1
DOIs
Publication statusPublished - 1 Jan 1991
Externally publishedYes

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