Cloning, sequencing, and enhanced expression of the dihydropteroate synthase gene of Escherichia coli MC4100.
Open accessAbstract
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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In E. coli and Plasmodium falciparum, nonsynonymous point mutations resulting in amino acid substitutions in DHPS can confer sulfa resistance.
“In other organisms, such as E. coli and Plasmodium falciparum, nonsynonymous point mutations resulting in amino acid substitutions in DHPS can confer sulfa resistance (14,15).”
From Dihydropteroate Synthase Gene Mutations in<i>Pneumocystis</i>and Sulfa Resistance (2004) · cites this paper as [14]
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Brooks, D. R., Wang, P., Read, M., Watkins, W. M., Sims, P. F. G., & Hyde, J. E. (1994) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1994.00397.x · Cited by 369 · see citing works
Helweg-Larsen, J., Benfield, T. L., Eugen-Olsen, J., Lundgren, J. D., & Lundgren, B. (1999) · The Lancet · doi:10.1016/s0140-6736(99)03320-6 · Cited by 274 · see citing works
Dihydropteroate synthase polymorphisms in Pneumocystis carinii.
Lane, B. R., Ast, J. C., Hossler, P. A., Mindell, D. P., Bartlett, M. S., Smith, J. W., & Meshnick, S. R. (1997) · Journal of Infectious Diseases · doi:10.1093/infdis/175.2.482 · Cited by 176 · see citing works
Dihydropteroate Synthase Gene Mutations in<i>Pneumocystis</i>and Sulfa Resistance
Huang, L., Crothers, K., Atzori, C., Benfield, T., Miller, R., Rabodonirina, M., & Helweg-Larsen, J. (2004) · Emerging Infectious Diseases · doi:10.3201/eid1010.030994 · Cited by 131 · see citing works
“coli and Plasmodium falciparum, nonsynonymous point mutations resulting in amino acid substitutions in DHPS can confer sulfa resistance (14,15).”
BackgroundPregnancy-associated Malaria, Challenges and Prospects in Sub-Saharan Africa
Aguzie, I. O. N. (2018) · Clinics in Mother and Child Health · doi:10.4172/2090-7214.1000282 · Cited by 13 · see citing works
- Authors
- Dallas, W. S., Gowen, J. E., Ray, P. H., Cox, M. J., & Dev, I. K.
- Affiliations
- Department of Molecular Genetics and Microbiology, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.
- Venue
- Journal of Bacteriology
- Publisher
- American Society for Microbiology
- Published
- 1992-09
- Volume
- 174
- Issue
- 18
- Pages
- 5961-5970
- ISSN
- 0021-9193 (ISSN-L)
- 1098-5530
- PMC
- PMC207134
- OpenAlex
- W1549970050
- Cited by
- 111
- References
- 0
- Citations by year
- 2026: 1 · 2024: 2 · 2023: 1 · 2019: 4 · 2018: 7 · 2017: 5 · 2016: 1 · 2014: 3 · 2012: 4
- FWCI
- 3.07
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Journal of Bacteriology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- dihydropteroate synthase, gene cloning, gene sequencing, T7 expression system, enzyme purification, protein crystallization, temperature-sensitive mutants, point mutations, p-aminobenzoic acid, sulfathiazole, folate biosynthesis, enzyme specific activity
- MeSH terms
- Amino Acid Sequence, Base Sequence, Cloning, Molecular, Dihydropteroate Synthase, DNA Mutational Analysis, Escherichia coli, Genetic Complementation Test, Molecular Sequence Data, Nucleic Acid Conformation, Recombinant Proteins, RNA, Messenger, Sequence Homology, Nucleic Acid, Gene Library, Gene Expression Regulation, Bacterial, Mutagenesis
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