Studies on Protein of the Occlusion-Derived Virus of Bombyx mori L. Nucleopolyhedrovirus Using Two-Dimensional Gel Electrophoresis and Mass Spectrometry

【Objective】The occlusion-derived virus (ODV) is produced when nucleocapsids retained in the nucleus are enveloped using virus-induced intranuclear membranes which appear to be derived from the nuclear membrane.It is specially adapted for primary infection of the host midgut epithelium. As such, the virion must contain the proteins essential for host range determination and initiation of infection. So knowledge of virion composition is a prerequisite for functional investigation.【Method】 In the present study, two-dimensional gel electrophoresis and mass spectrometry techniques were used to identify the proteins present within or associated with the occlusion-derived virus (ODV) virion, the separated proteins were visualized by Coomassie-Brilliant-Blue-G-250 staining. All protein spots were identified by MS.【Result】The separated proteins revealed 70 spots for the sample. Among them, most proteins were concentrated in pI 5-9, which reached 61% of the total protein spots. The results of this study suggest that 20 proteins were identified with confidence. Interestingly, the result showed that 21 spots of GP41 and 9 spots of VP39 were identified from the gel, respectively. 【Conclusion】It is feasible to extract ODV protein by a combination of ultracentrifugation and PCR technique; two-dimensional gel electrophoresis combined with mass spectrometry is a rapid and effective way of studying virus proteomics.