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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
5
|
| pubmed:dateCreated |
1996-4-17
|
| pubmed:abstractText |
This study describes the use and the advantages of the green fluorescent protein (GFP) as a reporter molecule for mycobacteria. The gfp gene from Aequorea victoria was placed under the control of the hsp60 promoter in the shuttle vector pGFM-11. The gfp expression in the recombinant Mycobacterium smegmatis and BCG was readily detected on agar plates by the development of an intense green fluorescence upon irradiation with long-wave u.v. light. In mycobacteria containing a pGFM-11 derivative that lacks the hsp60 promoter, no fluorescence was observed. However, this plasmid was successfully used as a promoter-probe vector to identify BCG promoters. The fluorescence emission of GFP in mycobacteria harbouring pGFM-11 and grown in liquid media could be quantified by spectrofluorimetry. This allowed for easy assessment of drug susceptibility. As GFP does not require the addition of substrates or co-factors, the green fluorescent bacilli could be directly observed within infected macrophages using fluorescence and laser confocal microscopy, or in tissue sections of infected mice. Finally, infected cells or free-living recombinant mycobacteria could also be analysed by flow cytometry. The GFP thus appears to be a convenient reporter for mycobacteria, allowing tracing of recombinant mycobacteria, isolation of promoters with interesting properties, in vivo drug testing and the development of new diagnostic tools.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antitubercular Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Chaperonin 60,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Green Fluorescent Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Luminescent Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
0950-382X
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
17
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
913-22
|
| pubmed:dateRevised |
2008-11-21
|
| pubmed:meshHeading |
pubmed-meshheading:8596440-Amino Acid Sequence,
pubmed-meshheading:8596440-Animals,
pubmed-meshheading:8596440-Antitubercular Agents,
pubmed-meshheading:8596440-Base Sequence,
pubmed-meshheading:8596440-Cell Line,
pubmed-meshheading:8596440-Chaperonin 60,
pubmed-meshheading:8596440-DNA Primers,
pubmed-meshheading:8596440-Genetic Vectors,
pubmed-meshheading:8596440-Green Fluorescent Proteins,
pubmed-meshheading:8596440-Luminescent Proteins,
pubmed-meshheading:8596440-Macrophages,
pubmed-meshheading:8596440-Mice,
pubmed-meshheading:8596440-Microbial Sensitivity Tests,
pubmed-meshheading:8596440-Microscopy, Confocal,
pubmed-meshheading:8596440-Microscopy, Fluorescence,
pubmed-meshheading:8596440-Molecular Sequence Data,
pubmed-meshheading:8596440-Mycobacterium,
pubmed-meshheading:8596440-Mycobacterium bovis,
pubmed-meshheading:8596440-Polymerase Chain Reaction,
pubmed-meshheading:8596440-Promoter Regions, Genetic,
pubmed-meshheading:8596440-Recombinant Proteins
|
| pubmed:year |
1995
|
| pubmed:articleTitle |
Green fluorescent protein as a new expression marker in mycobacteria.
|
| pubmed:affiliation |
Laboratoire de Microbiologie Génétique et Moléculaire, INSERM CJF9109, Institut Pasteur de Lille, France.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|