Mutants of Agrobacterium tumefaciens with elevated vir gene expression

Gregory J. Pazour, N. Ta Christopher, Anath Das

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28 Scopus citations

Abstract

Expression of Agrobacterium tumefaciens virulence (vir) genes requires virA, virG, and a plant-derived inducing compound such as acetosyringone. To identify the critical functional domains of virA and virG, a mutational approach was used. Agrobacterium A136 harboring plasmid pGP159, which contains virA, virG, and a reporter virB:lacZ gene fusion, was mutagenized with UV light or nitrosoguanidine. Survivors that formed blue colonies on a plate containing 5-bromo-4-chloro-3-indolyl β-n-galactoside were isolated and analyzed. Quantification of β-galactosidase activity in liquid assays identified nine mutant strains. By plasmid reconstruction and other procedures, all mutations mapped to the virA locus. These mutations caused an 11- to 560-fold increase in the vegetative level of virB:lacZ reporter gene expression. DNA sequence analysis showed that the mutations are located in four regions of VirA: transmembrane domain one, the active site, a glycine-rich region with homology to ATP-binding sites, and a region at the C terminus that has homology to the N terminus of VirG.

Original languageEnglish (US)
Pages (from-to)6941-6945
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number16
DOIs
StatePublished - 1991

Keywords

  • Mutagenesis
  • Vir gene regulation
  • VirA mutants

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