Molecular and General Genetics MGG

, Volume 256, Issue 1, pp 84–87 | Cite as

Single-step conjugative cloning of bacterial gene fusions involved in microbe-host interactions

  • P. B. Rainey
  • D. M. Heithoff
  • M. J. Mahan
SHORT COMMUNICATION

Abstract

In vivo expression technology (IVET) is a genetic strategy for isolating genes expressed in vivo. In order to fully exploit this technology, it is necessary to analyse large numbers of IVET-generated gene fusions, which must be recovered from the chromosome of host bacteria. In bacteria for which transductional methods are not available, the recovery of integrated fusion plasmids is problematic and currently limits broad application of IVET. We describe a rapid, single-step, triparental conjugative approach for recovering chromosomally integrated fusion plasmids from both Pseudomonas fluorescens and Salmonella typhimurium. This simple and broadly applicable conjugative cloning system extends the utility of the IVET approach to clinically and agronomically relevant microbes and may be employed to recover non-replicating and integrated plasmids in other systems.

Key words In vivo expression technology (IVET) Pseudomonas Salmonella Conjugation Retrotransfer 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • P. B. Rainey
    • 1
  • D. M. Heithoff
    • 2
  • M. J. Mahan
    • 2
  1. 1.Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK Fax: +44-1865-275074; e-mail: prainey@worf.molbiol.ox.ac.ukGB
  2. 2.Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA 93106, USAUS

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