Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles

Abstract

Objectives: In this paper, we report the expression and assembly of VLP in cells of E. coli expressing the CMVCP gene. Materials and Methods: In this study, the CMV-CP gene was released from a previously prepared cloning vector. Then, the CMV-CP was ligated into the expression vector. Sequencing was done by Marcrogen, Inc. (South Korea). A recombinant plasmid was transferred to E.coli isolate Rosetta. After inducing by isopropyl thiogalactosides, the molecular weight of the expressed protein was determined by SDS-PAGE. The extraction of proteins was done by NATURE method to see the possible presence of CMV-like particles. Results: CMV-CP was detected by Western blotting by a CMV specific polyclonal antibody and conjugate. The protein extracted from the CP producing clone was studied under a JEOL 100-CXII transmission electron microscope with 100000× magnification at an acceleration voltage of 100 kV. Conclusions: The results showed that the CP gene was expressed in the prokaryotic system successfully and was assembled into the CMVlike particle.