Materials and Methods: Here, we use PacBio and Illumina sequencing to reveal the complete genome sequence of H pylori B128 7.13. Furthermore, we describe a system to generate markerless and scarless mutations on the H pylori chromosome using the counter‐selection marker, galactokinase from Escherichia coli.
Results: We show that this mutagenesis strategy can be used to generate in‐frame insertions, gene deletions, and multiple independent mutations in B128 7.13. Using the closed genome as a reference, we also report the absence of second site chromosomal mutations and/or rearrangements in our mutagenized strains. We compare the genome sequence of H pylori B128 7.13 with a closely related strain, H pylori B8, and reveal one notable region of difference, which is a 1430 bp insertion encoding a H pylori‐specific DUF874 family protein of unknown function.
Conclusions: This article reports the closed genome of the important H pylori B128 7.13 strain and a mutagenesis method that can be adopted by researchers as an alternative strategy to generate isogenic mutants of H pylori in order to further our understanding of this bacterium.
Bibliographical note© 2019. The Authors. Helicobacter Published by John Wiley & Sons Ltd.
- gene mutation
- Helicobacter pylori