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Cloning and functional expression of a mungbean defensin VrD1 in Pichia pastoris
Authors:Chen Ji-Jr  Chen Gan-Hong  Hsu Hui-Ching  Li Shin-Shing  Chen Ching-San
Institution:Graduate Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung 202, Taiwan.
Abstract:It was shown previously that a bacterially expressed mungbean defensin VrCRP exhibited both antifungal and insecticidal activities. To isolate this protein in a large quantity for its characterization, the defensin cDNA was expressed in Pichia pastoris and the recombinant defensin (rVrD1) was purified. The recombinant VrD1 was shown to inhibit the growth of fungi such as Fusarium oxysporum, Pyricularia oryza, Rhizoctonia solani, and Trichophyton rubrum and development of bruchid larva. The protein also inhibits in vitro protein synthesis. These biological activities are similar to that of the bacterially expressed defensin. Functional expression of VrD1 in Pichia pastoris provides a highly feasible system to study the structure-function relationship of VrD1 using the mutagenesis approach.
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