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Helicobacter pylori with separate β‐ and β′‐subunits of RNA polymerase is viable and can colonize conventional mice
Author(s) -
Raudonikiene Ausra,
Zakharova Natalya,
Su Wan Wen,
Jeong JinYong,
Bryden Louis,
Hoffman Paul S.,
Berg Douglas E.,
Severinov Konstantin
Publication year - 1999
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.1999.01336.x
Subject(s) - rpob , biology , rna polymerase , polymerase , gene , helicobacter pylori , microbiology and biotechnology , polymerase chain reaction , genetics , open reading frame , rna , 16s ribosomal rna , peptide sequence
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 electrotransformation. 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.