Molecular analysis of urease genes from a newly identified uncultured species of Helicobacter

Jay V Solnick, J. O'Rourke, A. Lee, L. S. Tompkins

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

'Gastrospirillum hominis' is an uncultured gastric spiral bacterium that has recently been shown by 16S rDNA sequence analysis to be a newly recognized species of Helicobacter that infects humans, and it has been provisionally designated 'Helicobacter heilmannii.' We used PCR to directly amplify the urease structural genes of 'H. heilmannii' from infected gastric tissue. DNA sequence analysis identified two open reading frames, ureA and ureB, which code for polypeptides with predicted molecular weights of 25,729 and 61,831, respectively. The urease subunit genes from 'H. heilmannii' were cloned and expressed in Escherichia coli. Western blot (immunoblot) analysis showed that antiserum directed against the ureA and ureB gene products from H. pylori was cross-reactive with the corresponding polypeptides from 'H. heilmannii.' Analysis of the derived amino acid sequences of 'H. heilmannii' UreA and UreB demonstrated that 'H. heilmannii' urease is more highly related to the urease from H. felis (found in the stomachs of cats and dogs) than to the urease from H. pylori. These data are consistent with 16S rDNA sequence analysis and suggest that 'H. heilmannii' is phylogenetically most closely related to H. felis.

Original languageEnglish (US)
Pages (from-to)1631-1638
Number of pages8
JournalInfection and Immunity
Volume62
Issue number5
StatePublished - 1994
Externally publishedYes

Fingerprint

Helicobacter heilmannii
Helicobacter
Urease
Genes
Felis
Urea
Stomach
Pylorus
Ribosomal DNA
Sequence Analysis
Western Blotting
Peptides
Protein Sequence Analysis
DNA Sequence Analysis
Open Reading Frames
Immune Sera
Cats
Molecular Weight
Dogs
Escherichia coli

ASJC Scopus subject areas

  • Immunology

Cite this

Molecular analysis of urease genes from a newly identified uncultured species of Helicobacter. / Solnick, Jay V; O'Rourke, J.; Lee, A.; Tompkins, L. S.

In: Infection and Immunity, Vol. 62, No. 5, 1994, p. 1631-1638.

Research output: Contribution to journalArticle

Solnick, Jay V ; O'Rourke, J. ; Lee, A. ; Tompkins, L. S. / Molecular analysis of urease genes from a newly identified uncultured species of Helicobacter. In: Infection and Immunity. 1994 ; Vol. 62, No. 5. pp. 1631-1638.
@article{9258ac1ba66f4157989e945625dca832,
title = "Molecular analysis of urease genes from a newly identified uncultured species of Helicobacter",
abstract = "'Gastrospirillum hominis' is an uncultured gastric spiral bacterium that has recently been shown by 16S rDNA sequence analysis to be a newly recognized species of Helicobacter that infects humans, and it has been provisionally designated 'Helicobacter heilmannii.' We used PCR to directly amplify the urease structural genes of 'H. heilmannii' from infected gastric tissue. DNA sequence analysis identified two open reading frames, ureA and ureB, which code for polypeptides with predicted molecular weights of 25,729 and 61,831, respectively. The urease subunit genes from 'H. heilmannii' were cloned and expressed in Escherichia coli. Western blot (immunoblot) analysis showed that antiserum directed against the ureA and ureB gene products from H. pylori was cross-reactive with the corresponding polypeptides from 'H. heilmannii.' Analysis of the derived amino acid sequences of 'H. heilmannii' UreA and UreB demonstrated that 'H. heilmannii' urease is more highly related to the urease from H. felis (found in the stomachs of cats and dogs) than to the urease from H. pylori. These data are consistent with 16S rDNA sequence analysis and suggest that 'H. heilmannii' is phylogenetically most closely related to H. felis.",
author = "Solnick, {Jay V} and J. O'Rourke and A. Lee and Tompkins, {L. S.}",
year = "1994",
language = "English (US)",
volume = "62",
pages = "1631--1638",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "5",

}

TY - JOUR

T1 - Molecular analysis of urease genes from a newly identified uncultured species of Helicobacter

AU - Solnick, Jay V

AU - O'Rourke, J.

AU - Lee, A.

AU - Tompkins, L. S.

PY - 1994

Y1 - 1994

N2 - 'Gastrospirillum hominis' is an uncultured gastric spiral bacterium that has recently been shown by 16S rDNA sequence analysis to be a newly recognized species of Helicobacter that infects humans, and it has been provisionally designated 'Helicobacter heilmannii.' We used PCR to directly amplify the urease structural genes of 'H. heilmannii' from infected gastric tissue. DNA sequence analysis identified two open reading frames, ureA and ureB, which code for polypeptides with predicted molecular weights of 25,729 and 61,831, respectively. The urease subunit genes from 'H. heilmannii' were cloned and expressed in Escherichia coli. Western blot (immunoblot) analysis showed that antiserum directed against the ureA and ureB gene products from H. pylori was cross-reactive with the corresponding polypeptides from 'H. heilmannii.' Analysis of the derived amino acid sequences of 'H. heilmannii' UreA and UreB demonstrated that 'H. heilmannii' urease is more highly related to the urease from H. felis (found in the stomachs of cats and dogs) than to the urease from H. pylori. These data are consistent with 16S rDNA sequence analysis and suggest that 'H. heilmannii' is phylogenetically most closely related to H. felis.

AB - 'Gastrospirillum hominis' is an uncultured gastric spiral bacterium that has recently been shown by 16S rDNA sequence analysis to be a newly recognized species of Helicobacter that infects humans, and it has been provisionally designated 'Helicobacter heilmannii.' We used PCR to directly amplify the urease structural genes of 'H. heilmannii' from infected gastric tissue. DNA sequence analysis identified two open reading frames, ureA and ureB, which code for polypeptides with predicted molecular weights of 25,729 and 61,831, respectively. The urease subunit genes from 'H. heilmannii' were cloned and expressed in Escherichia coli. Western blot (immunoblot) analysis showed that antiserum directed against the ureA and ureB gene products from H. pylori was cross-reactive with the corresponding polypeptides from 'H. heilmannii.' Analysis of the derived amino acid sequences of 'H. heilmannii' UreA and UreB demonstrated that 'H. heilmannii' urease is more highly related to the urease from H. felis (found in the stomachs of cats and dogs) than to the urease from H. pylori. These data are consistent with 16S rDNA sequence analysis and suggest that 'H. heilmannii' is phylogenetically most closely related to H. felis.

UR - http://www.scopus.com/inward/record.url?scp=0028215785&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028215785&partnerID=8YFLogxK

M3 - Article

C2 - 8168924

AN - SCOPUS:0028215785

VL - 62

SP - 1631

EP - 1638

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 5

ER -