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PMID: 2982791 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Identification and genetic analysis of an Agrobacterium tumefaciens chromosomal virulence region.

Journal of bacteriology ·Vol. 161 ·No. 3 ·1985-03-00 ·Pages 850-60

Douglas CJ, Staneloni RJ, Rubin RA, Nester EW

Abstract

A genetic analysis of Agrobacterium tumefaciens chromosomal functions required for virulence was undertaken. Large Tn5-containing cosmid clones were isolated from DNA of avirulent A. tumefaciens mutants having chromosomal Tn5 insertions and exhibiting defective attachment to plant cells. The clones from several different mutants each contained overlapping segments of a 30-kilobase A. tumefaciens chromosomal region, which were physically mapped. All chromosomal Tn5 insertions leading to the avirulent, attachment-defective phenotype were localized within an 11-kilobase portion of this chromosomal virulence region. Transposon Tn3::HoHo1 (Tn3 containing lacZ) was used to simultaneously mutagenize and create lac fusions within the virulence region. This analysis demonstrated the presence of two distinct chromosomal virulence loci, which were 1.5 and 5 kilobases long; transposon insertions into these loci led to avirulence and defective attachment. The beta-galactosidase activity associated with various Tn3::HoHo1-created lac fusions indicated that the loci are transcribed in opposite directions, and complementation studies suggested that each locus consists of a single transcriptional unit. A cosmid clone of the chromosomal virulence region containing a lac fusion in the extreme 3' portion of the 5-kilobase locus was used to demonstrate that expression of this region is dependent on the presence of sequences in the 5' portion of the locus, confirming its operon-like nature.

MeSH Terms
Chromosomes, Bacterial/ultrastructure DNA Restriction Enzymes DNA Transposable Elements DNA, Bacterial/genetics Genes, Bacterial Genetic Complementation Test Mutation Plasmids Rhizobium/genetics,pathogenicity Transcription, Genetic
Chemicals
DNA Transposable Elements DNA, Bacterial DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Douglas C J
Staneloni R J
Rubin R A
Nester E W
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41 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.161.3.850-860.1985
Published
1985-03-00
Pages
850-60
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC214975
Subset
IM
Grants
NIGMS NIH HHS · 5 R01 GM32618-12 · United States
External Links
PubMed source DOI: 10.1128/jb.161.3.850-860.1985 PMC Full Text View journal details
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