Helicobacter pylori with separate β- and β'-subunits of RNA polymerase is viable and can colonize conventional mice

Ausra Raudonikiene, Natalya Zakharova, Wan Wen Su, Jin Yong Jeong, Louis Bryden, Paul S. Hoffman, Douglas E. Berg, Konstantin Severinov

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The genes encoding the β- and β'-subunits of RNA polymerase (rpoB and rpoC respectively) are fused as one continuous open reading frame in Helicobacter pylori and in other members of this genus, but are separate in other bacterial taxonomic groups, including the closely related genus Campylobacter. To test whether this β-β' tethering is essential, we used polymerase chain reaction-based cloning to separate the rpoB and rpoC moieties of the H. pylori rpoB-rpoC fusion gene with a non-polar chloramphenicol resistance cassette containing a new translational start, and introduced this construct into H. pylori by electro-transformation. H. pylori containing these separated rpoB and rpoC genes in place of the native fusion gene produced non-tethered β and β' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/6 mice. Thus, the extraordinary β-β' tethering is not essential for H. pylori viability and gastric colonization.

Original languageEnglish (US)
Pages (from-to)131-138
Number of pages8
JournalMolecular microbiology
Volume32
Issue number1
DOIs
StatePublished - Apr 19 1999

Fingerprint

DNA-Directed RNA Polymerases
Helicobacter pylori
Gene Fusion
Chloramphenicol Resistance
Campylobacter
Inbred C57BL Mouse
Open Reading Frames
Genes
Organism Cloning
Stomach
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Microbiology

Cite this

Raudonikiene, Ausra ; Zakharova, Natalya ; Su, Wan Wen ; Jeong, Jin Yong ; Bryden, Louis ; Hoffman, Paul S. ; Berg, Douglas E. ; Severinov, Konstantin. / Helicobacter pylori with separate β- and β'-subunits of RNA polymerase is viable and can colonize conventional mice. In: Molecular microbiology. 1999 ; Vol. 32, No. 1. pp. 131-138.
@article{6ea80548dad84d6ea3abeb1403a6599a,
title = "Helicobacter pylori with separate β- and β'-subunits of RNA polymerase is viable and can colonize conventional mice",
abstract = "The genes encoding the β- and β'-subunits of RNA polymerase (rpoB and rpoC respectively) are fused as one continuous open reading frame in Helicobacter pylori and in other members of this genus, but are separate in other bacterial taxonomic groups, including the closely related genus Campylobacter. To test whether this β-β' tethering is essential, we used polymerase chain reaction-based cloning to separate the rpoB and rpoC moieties of the H. pylori rpoB-rpoC fusion gene with a non-polar chloramphenicol resistance cassette containing a new translational start, and introduced this construct into H. pylori by electro-transformation. H. pylori containing these separated rpoB and rpoC genes in place of the native fusion gene produced non-tethered β and β' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/6 mice. Thus, the extraordinary β-β' tethering is not essential for H. pylori viability and gastric colonization.",
author = "Ausra Raudonikiene and Natalya Zakharova and Su, {Wan Wen} and Jeong, {Jin Yong} and Louis Bryden and Hoffman, {Paul S.} and Berg, {Douglas E.} and Konstantin Severinov",
year = "1999",
month = "4",
day = "19",
doi = "https://doi.org/10.1046/j.1365-2958.1999.01336.x",
language = "English (US)",
volume = "32",
pages = "131--138",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "1",

}

Helicobacter pylori with separate β- and β'-subunits of RNA polymerase is viable and can colonize conventional mice. / Raudonikiene, Ausra; Zakharova, Natalya; Su, Wan Wen; Jeong, Jin Yong; Bryden, Louis; Hoffman, Paul S.; Berg, Douglas E.; Severinov, Konstantin.

In: Molecular microbiology, Vol. 32, No. 1, 19.04.1999, p. 131-138.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Helicobacter pylori with separate β- and β'-subunits of RNA polymerase is viable and can colonize conventional mice

AU - Raudonikiene, Ausra

AU - Zakharova, Natalya

AU - Su, Wan Wen

AU - Jeong, Jin Yong

AU - Bryden, Louis

AU - Hoffman, Paul S.

AU - Berg, Douglas E.

AU - Severinov, Konstantin

PY - 1999/4/19

Y1 - 1999/4/19

N2 - The genes encoding the β- and β'-subunits of RNA polymerase (rpoB and rpoC respectively) are fused as one continuous open reading frame in Helicobacter pylori and in other members of this genus, but are separate in other bacterial taxonomic groups, including the closely related genus Campylobacter. To test whether this β-β' tethering is essential, we used polymerase chain reaction-based cloning to separate the rpoB and rpoC moieties of the H. pylori rpoB-rpoC fusion gene with a non-polar chloramphenicol resistance cassette containing a new translational start, and introduced this construct into H. pylori by electro-transformation. H. pylori containing these separated rpoB and rpoC genes in place of the native fusion gene produced non-tethered β and β' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/6 mice. Thus, the extraordinary β-β' tethering is not essential for H. pylori viability and gastric colonization.

AB - The genes encoding the β- and β'-subunits of RNA polymerase (rpoB and rpoC respectively) are fused as one continuous open reading frame in Helicobacter pylori and in other members of this genus, but are separate in other bacterial taxonomic groups, including the closely related genus Campylobacter. To test whether this β-β' tethering is essential, we used polymerase chain reaction-based cloning to separate the rpoB and rpoC moieties of the H. pylori rpoB-rpoC fusion gene with a non-polar chloramphenicol resistance cassette containing a new translational start, and introduced this construct into H. pylori by electro-transformation. H. pylori containing these separated rpoB and rpoC genes in place of the native fusion gene produced non-tethered β and β' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/6 mice. Thus, the extraordinary β-β' tethering is not essential for H. pylori viability and gastric colonization.

UR - http://www.scopus.com/inward/record.url?scp=0032939352&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032939352&partnerID=8YFLogxK

U2 - https://doi.org/10.1046/j.1365-2958.1999.01336.x

DO - https://doi.org/10.1046/j.1365-2958.1999.01336.x

M3 - Article

C2 - 10216866

VL - 32

SP - 131

EP - 138

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 1

ER -