rdf:type |
|
lifeskim:mentions |
|
pubmed:issue |
2
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pubmed:dateCreated |
1999-1-28
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pubmed:databankReference |
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pubmed:abstractText |
A cDNA clone encoding a general odorant-binding protein (GOBP2) was isolated from antennal RNA of Mamestra brassicae by reverse transcription-PCR (RT-PCR) and RACE-PCR. The cDNA encoding the GOBP2 was further used for bacterial expression. Most of the recombinant GOBP2 (>90%) was found to be insoluble. Purification under denaturing conditions consisted of solubilisation of inclusion bodies, affinity chromatography, refolding and gel filtration. The refolded rGOBP2 was cross-reactive with a serum raised against the GOBP2 of the Lepidoptera Antheraea polyphemus. The purified refolded rGOBP2 was further characterised by native PAGE, IEF, N-terminal sequencing, and two-dimensional NMR. A functional characterisation of the rGOBP2 was carried out by testing its ability to bind pheromone compounds. The yields of production and purification fulfil the requirements of structural studies.
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pubmed:language |
eng
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pubmed:journal |
|
pubmed:citationSubset |
IM
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pubmed:chemical |
|
pubmed:status |
MEDLINE
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pubmed:month |
Dec
|
pubmed:issn |
0014-2956
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pubmed:author |
|
pubmed:issnType |
Print
|
pubmed:day |
1
|
pubmed:volume |
258
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
768-74
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pubmed:dateRevised |
2007-7-23
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pubmed:meshHeading |
pubmed-meshheading:9874246-Amino Acid Sequence,
pubmed-meshheading:9874246-Animals,
pubmed-meshheading:9874246-Base Sequence,
pubmed-meshheading:9874246-Blotting, Southern,
pubmed-meshheading:9874246-Cloning, Molecular,
pubmed-meshheading:9874246-Insects,
pubmed-meshheading:9874246-Magnetic Resonance Spectroscopy,
pubmed-meshheading:9874246-Molecular Sequence Data,
pubmed-meshheading:9874246-Pheromones,
pubmed-meshheading:9874246-Protein Binding,
pubmed-meshheading:9874246-Protein Folding,
pubmed-meshheading:9874246-RNA, Messenger,
pubmed-meshheading:9874246-Receptors, Odorant,
pubmed-meshheading:9874246-Recombinant Proteins,
pubmed-meshheading:9874246-Sequence Analysis, DNA,
pubmed-meshheading:9874246-Sequence Homology, Amino Acid
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pubmed:year |
1998
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pubmed:articleTitle |
Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae.
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pubmed:affiliation |
INRA, Unité de Phytopharmacie et des Médiateurs Chimiques, Versailles, France.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|