التفاصيل البيبلوغرافية
| العنوان: |
Expression of Fragaria ananassa Osmotin-like Protein (FaOLP2) in E. coli: Purification and Antifungal Activity. |
| المؤلفون: |
LI, Bai, LI, Sufang, LI, Huandi, SUN, Junwei, ZOU, Keqin |
| المصدر: |
Agricultural Biotechnology (2164-4993); Dec2013, Vol. 2 Issue 6, p7-11, 5p |
| مصطلحات موضوعية: |
STRAWBERRIES, OSMOTIC potential of plants, RECOMBINANT proteins, ESCHERICHIA coli, ANTIFUNGAL agents, GENE expression in plants |
| مستخلص: |
[Objective] The FaOLPl (Fragaria ananassa osmotin-like protein) is a functional homolog of PR5-like protein. This study was undertaken to produce recombinant FaOLP2 and to identify its antifungal activity. [Method] The ORF of FaOLP2 (accession number DQ325524) was cloned into pET22b vector to construct the pET22b-FaOLP2 plasmid. The recombinant mature FaOLP2 was expressed in E. coli Rosetta-gami B (DE3) by inducing with 1 mmol/L IPTG and found exclusively in insoluble inclusion bodies. As FaOLP2 requires the correct formation of eight disulfide bonds, but there were no obvious effect to correctly form these by expression at different temperatures and high osmotic pressure (supplemented Betaineand and D-Sorbitol), we used an in vitro method to refold E. coli expressed FaOLP2 by gradually elution using reduced;oxidized gluthatione redox buffer, followed by 8 mol/L urea solubilized His6-tagged mature FaOLP2 protein, which was affinity-purified by an immobilized-metal (Ni+2) affinity chromatography (IMAC) column. [Result] This method generated biologically active conformations of the recombinant mature FaOLP2 that displayed antifungal activity against Ustilaginoides virens, a plant pathogenic fungus, which causes rice false smut. [Conclusion] This study laid the foundation for further biotechnological application of the novel protein. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
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