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A family of promoter probe vectors incorporating autofluorescent and chromogenic reporter proteins for studying gene expression in gram-negative bacteria

  • Arthur Hosie
    ,
  • R. Karunakaran
    ,
  • T.H. Mauchline
    ,
  • Philip S. Poole
  • University of Reading
Research Output: Contribution to journal Article Peer-review

Abstract

A series of promoter probe vectors for use in Gram-negative bacteria has been made in two broad-host-range vectors, pOT (pBBR replicon) and pJP2 (incP replicon). Reporter fusions can be made to gfpUV, gfpmut3.1, unstable gfpmut3.1 variants (LAA, LVA, AAV and ASV), gfp+, dsRed2, dsRedT.3, dsRedT.4, mRFP1, gusA or lacZ. The two vector families, pOT and pJP2, are compatible with one another and share the same polylinker for facile interchange of promoter regions. Vectors based on pJP2 have the advantage of being ultra-stable in the environment due to the presence of the parABCDE genes. As a confirmation of their usefulness, the dicarboxylic acid transport system promoter (dctAp) was cloned into a pOT (pRU1097)- and a pJP2 (pRU1156)-based vector and shown to be expressed by Rhizobium leguminosarum in infection threads of vetch. This indicates the presence of dicarboxylates at the earliest stages of nodule formation.

Publication Information

Output type

Research Output: Contribution to journal Article Peer-review

Original language

English

Pages from-to (Number of pages)

Pages 3249-3256

Journal (Volume, Issue Number)

Microbiology (United Kingdom) (Volume 151, Issue 10)

Publication milestones

  • Published - 01/01/2005

Publication status

Published - 01/01/2005

ISSN

1350-0872

External Publication IDs

  • handle.net: 10547/293938
  • Scopus: 27144488219

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