A practical guide to recombineering in Photorhabdus and Xenorhabdus

Research output: Contribution to book/conference proceedings/anthology/reportChapter in book/anthology/reportContributedpeer-review

Contributors

  • Jia Yin - (Author)
  • Hailong Wang - (Author)
  • Ruijuan Li - (Author)
  • Vinothkannan Ravichandran - (Author)
  • Xiaoying Bian - (Author)
  • Aiying Li - (Author)
  • Qiang Tu - (Author)
  • A. Francis Stewart - , Chair of Applied Genomics (Author)
  • Jun Fu - , TUD Dresden University of Technology (Author)
  • Youming Zhang - (Author)

Abstract

Fluent genetic manipulation of prokaryote genomes is still limited to only a few commonly used hosts. Ideally the advanced technologies available for cloning into recombinant Escherichia coli should also be applicable in other prokaryotes. In particular, ‘recombineering’ is mediated by the lambda Red operon that permits fluent and precise engineering of the E. coli genome and associated recombinant DNA. The major limitation is that host-specific phage-derived recombination systems are also required in more distant species. Recently, an endogenous Red-like operon Pluγβα has been reported to be effective in both Photorhabdus and Xenorhabdus bacteria. The Pluγβα recombineering system is based on three host-specific phage proteins from Photorhabdus luminescens, Plu2935, Plu2936, and Plu2934, which are functional analogs of Redβ, Redα, and Redγ, respectively. In this chapter, we provide a comprehensive and up-to-date method for P. luminescens and Xenorhabdus stockiae genome engineering via the Pluγβα recombineering system. In order to facilitate the rapid construction of knock-in vectors, recET-mediated recombineering is incorporated in the pipeline. Concerted recET system in E. coli with Pluγβα system in Photorhabdus and Xenorhabdus could promote reverse genetics, functional genomics, and bioprospecting research for these two genera.

Details

Original languageEnglish
Title of host publicationCurrent Topics in Microbiology and Immunology
PublisherSpringer Verlag
Pages195-213
Number of pages19
Publication statusPublished - 2017
Peer-reviewedYes

Publication series

SeriesCurrent topics in microbiology and immunology : CTMI = Ergebnisse der Mikrobiologie und Immunitätsforschung
Volume402
ISSN0070-217X

External IDs

PubMed 28154939
ORCID /0000-0002-4754-1707/work/142248104