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PMID: 10216866 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Molecular microbiology ·Vol. 32 ·No. 1 ·1999-04-00 ·Pages 131-8

Raudonikiene A, Zakharova N, Su WW, Jeong JY, Bryden L, Hoffman PS, Berg DE, Severinov K

Abstract

The genes encoding the beta- and beta'-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 beta-beta' 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 beta and beta' RNAP subunits, grew well in culture and colonized and proliferated well in conventional C57BL/6 mice. Thus, the extraordinary beta-beta' tethering is not essential for H. pylori viability and gastric colonization.

MeSH Terms
Animals Chloramphenicol O-Acetyltransferase/metabolism Cloning, Molecular DNA-Directed RNA Polymerases/genetics,metabolism Electrophoresis, Polyacrylamide Gel Electroporation Female Helicobacter pylori/enzymology,pathogenicity Mice/microbiology Mice, Inbred C57BL Models, Genetic Mutagenesis, Insertional Plant Proteins/genetics Polymerase Chain Reaction Recombinant Fusion Proteins Stomach/microbiology Time Factors Transformation, Genetic
Chemicals
Plant Proteins Recombinant Fusion Proteins RpoB protein, Oryza sativa Chloramphenicol O-Acetyltransferase beta' subunit of RNA polymerase DNA-Directed RNA Polymerases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Raudonikiene A
Department of Molecular Microbiology, Washington University School of Medicine, St Louis 63110, MO, USA.
Zakharova N
Su W W
Jeong J Y
Bryden L
Hoffman P S
Berg D E
Severinov K
Article Info
Journal
Molecular microbiology
Abbr.
Mol Microbiol
ISSN
0950-382X
Published
1999-04-00
Pages
131-8
Language
English
Region
England
NLM ID
8712028
Subset
IM
Grants
NIAID NIH HHS · AI 38166 · United States
NIDDK NIH HHS · DK48029 · United States
NHGRI NIH HHS · HG00820 · United States
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