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PMID: 1373718 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Tracking genetically engineered bacteria: monoclonal antibodies against surface determinants of the soil bacterium Pseudomonas putida 2440.

Journal of bacteriology ·Vol. 174 ·No. 9 ·1992-05-00 ·Pages 2978-85

Ramos-González MI, Ruiz-Cabello F, Brettar I, Garrido F, Ramos JL

Abstract

Assessment of potential risks involved in the release of genetically engineered microorganisms is facilitated by the availability of monoclonal antibodies (MAbs), a tool potentially able to monitor specific organisms. We raised a bank of MAbs against the soil bacterium Pseudomonas putida 2440, which is a host for modified TOL plasmids and other recombinant plasmids. Three MAbs, 7.3B, 7.4D, and 7.5D, were highly specific and recognized only P. putida bacteria. Furthermore, we developed a semiquantitative dot blot assay that allowed us to detect as few as 100 cells per spot. A 40-kDa cell surface protein was the target for MAbs 7.4D and 7.5D. Detection of the cell antigen depended on the bacterial growth phase and culture medium. The O antigen of lipopolysaccharide seems to be the target for MAb 7.3B, and its in vivo detection was independent of the bacterial growth phase and culture medium. MAb 7.3B was used successfully to track P. putida (pWW0) released in unsterile lake mesocosms.

MeSH Terms
Antibodies, Monoclonal/immunology,isolation & purification Antigens, Bacterial/immunology,isolation & purification Bacterial Outer Membrane Proteins/immunology,isolation & purification Blotting, Western Environmental Microbiology Enzyme-Linked Immunosorbent Assay Genetic Engineering/methods O Antigens Polysaccharides, Bacterial/immunology,isolation & purification Pseudomonas putida/genetics,immunology,isolation & purification Soil Microbiology Water Microbiology
Chemicals
Antibodies, Monoclonal Antigens, Bacterial Bacterial Outer Membrane Proteins O Antigens Polysaccharides, Bacterial
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ramos-González M I
Consejo Superior de Investigaciones Científicas, Departamento de Bioquímica, Vegetal, Granada, Spain.
Ruiz-Cabello F
Brettar I
Garrido F
Ramos J L
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.174.9.2978-2985.1992
Published
1992-05-00
Pages
2978-85
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205952
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.174.9.2978-2985.1992 PMC Full Text View journal details
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