التفاصيل البيبلوغرافية
| العنوان: |
Molecular cloning and bacterial expression of a general odorant-binding protein from the cabbage armyworm Mamestra brassicae. |
| المؤلفون: |
Maibèche-Coisne, Martine1, Longhi, Sonia2, Jacquin-Joly, Emmanuelle1, Brunel, Carole2, Egloff, Marie-Pierre2, Gastinel, Louis2, Cambillau, Christian2, Tegoni, Mariella2, Nagnan-Le Meillour, Patricia1 |
| المصدر: |
European Journal of Biochemistry. Dec98 Part 1, Vol. 258 Issue 2, p768-774. 7p. 4 Black and White Photographs, 1 Diagram, 1 Graph. |
| مصطلحات موضوعية: |
*CARRIER proteins, *MOLECULAR cloning, *ARMYWORMS |
| مستخلص: |
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. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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