Molecular cloning and characterization of the aroD gene encoding 3-dehydroquinase from Salmonella typhi.
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.
Servos
S
S
Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, UK.
Chatfield
S
S
Hone
D
D
Levine
M
M
Dimitriadis
G
G
Pickard
D
D
Dougan
G
G
Fairweather
N
N
Charles
I
I
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006836:Hydro-Lyases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012485:Salmonella typhi / Q000201:enzymology / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D006836:Hydro-Lyases; C020000:3-dehydroquinate dehydratase