Nucleotide sequence of the gyrA gene and characterization of ciprofloxacin-resistant mutants of Helicobacter pylori.

Abstract:

:PCR was used to amplify a 238-bp region from Helicobacter pylori which corresponded to the quinolone resistance-determining region in Escherichia coli. The gyrA gene of H. pylori was cloned and sequenced. An open reading frame of 2,478 nucleotides coded for a polypeptide of 826 amino acids with a calculated molecular mass of 92,508 Da. The amino acid sequence showed an overall 52% identity with other bacterial gyrA genes but was most closely related to the gyrA subunit of Campylobacter jejuni (76.5% identity). Sequencing of the amplification product from ciprofloxacin-resistant mutants of H. pylori revealed four classes of mutations with substitutions at amino acid 87 (Asn-->Lys), amino acid 88 (Ala-->Val), and amino acid 91 (Asp-->Gly, -->Asn, or -->Tyr) and a double substitution at amino acids 91 and 97 (Ala-->Val). Ciprofloxacin-susceptible strains of H. pylori could be transformed to ciprofloxacin resistance by using the amplified fragment from resistant strains as donor DNA. Of the 11 ciprofloxacin-resistant mutants examined, only one did not have an alteration within the quinolone resistance-determining region, suggesting that, in H. pylori, resistance to quinolones is primarily a result of alterations in gyrA.

authors

Moore RA,Beckthold B,Wong S,Kureishi A,Bryan LE

doi

10.1128/aac.39.1.107

subject

Has Abstract

pub_date

1995-01-01 00:00:00

pages

107-11

issue

1

eissn

0066-4804

issn

1098-6596

journal_volume

39

pub_type

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