Molecular cloning and characterization of the aroD gene encoding 3-dehydroquinase from Salmonella typhi |
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Authors: | S Servos S Chatfield D Hone M Levine G Dimitriadis D Pickard G Dougan N Fairweather I Charles |
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Affiliation: | Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, UK. |
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Abstract: | The aroD gene from Salmonella typhi, encoding 5-dehydroquinate hydrolyase (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 Mr of 27706. 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-related with its S. typhi counterpart. |
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