Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae.
Eur J Biochem
; 258(2): 768-74, 1998 Dec 01.
Article
em En
| MEDLINE
| ID: mdl-9874246
ABSTRACT
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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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Receptores Odorantes
/
Insetos
Limite:
Animals
Idioma:
En
Revista:
Eur J Biochem
Ano de publicação:
1998
Tipo de documento:
Article
País de afiliação:
França