| Title: | Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae |
| Author(s): | Maibeche-Coisne M; Longhi S; Jacquin-Joly E; Brunel C; Egloff MP; Gastinel L; Cambillau C; Tegoni M; Nagnan-Le Meillour P; |
| Address: | "INRA, Unite de Phytopharmacie et des Mediateurs Chimiques, Versailles, France" |
| DOI: | 10.1046/j.1432-1327.1998.2580768.x |
| ISSN/ISBN: | 0014-2956 (Print) 0014-2956 (Linking) |
| 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" |
| Keywords: | "Amino Acid Sequence Animals Base Sequence Blotting, Southern Cloning, Molecular Insecta/*chemistry Magnetic Resonance Spectroscopy Molecular Sequence Data Pheromones/metabolism Protein Binding/physiology Protein Folding RNA, Messenger/genetics Receptors, ;" |
| Notes: | "MedlineMaibeche-Coisne, M Longhi, S Jacquin-Joly, E Brunel, C Egloff, M P Gastinel, L Cambillau, C Tegoni, M Nagnan-Le Meillour, P eng Research Support, Non-U.S. Gov't England 1999/01/05 Eur J Biochem. 1998 Dec 1; 258(2):768-74. doi: 10.1046/j.1432-1327.1998.2580768.x" |
