Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae
| العنوان: | Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae |
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| المؤلفون: | Mariella Tegoni, Christian Cambillau, Marie-Pierre Egloff, Sonia Longhi, Martine Maïbèche-Coisne, Emmanuelle Jacquin-Joly, Carole Brunel, Patricia Nagnan-Le Meillour, Louis Noël Gastinel |
| المساهمون: | Unité de phytopharmacie et médiateurs chimiques, Institut National de la Recherche Agronomique (INRA), ProdInra, Migration |
| المصدر: | European Journal of Biochemistry European Journal of Biochemistry, Wiley, 1998, 258, pp.768-774 |
| سنة النشر: | 1999 |
| مصطلحات موضوعية: | Protein Folding, Insecta, Magnetic Resonance Spectroscopy, Sequence analysis, Molecular Sequence Data, Molecular cloning, Biology, Receptors, Odorant, Biochemistry, Inclusion bodies, Pheromones, law.invention, Antheraea polyphemus, 03 medical and health sciences, 0302 clinical medicine, Affinity chromatography, law, Complementary DNA, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, RNA, Messenger, Cloning, Molecular, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Base Sequence, Sequence Homology, Amino Acid, RNA, Sequence Analysis, DNA, biology.organism_classification, Molecular biology, Recombinant Proteins, Blotting, Southern, Recombinant DNA, 030217 neurology & neurosurgery, Protein Binding |
| الوصف: | 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. |
| تدمد: | 0014-2956 1432-1327 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::419b9230e7f7c56c0587a104cde9388dTest https://pubmed.ncbi.nlm.nih.gov/9874246Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....419b9230e7f7c56c0587a104cde9388d |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00142956 14321327 |
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