Author/Authors :
Rostami، Afshin نويسنده Plant Protection Department, University of Zanjan, Zanjan, IR Iran , , Sokhandan Bashir، Nemat نويسنده Plant Protection Department, University of Tabriz, P.O. Box 345, Tabriz, I.R. Iran , , Pirniakan، Parviz نويسنده Department of Agriculture, Payame Noor University, IR Iran , , Masoudi، Nahid نويسنده Department of Plant Protection, University of Guilan, Rasht, IR Iran ,
Abstract :
Cucumber mosaic virus (CMV) has isometric particles with a diameter of about 28–29 nm. A useful method to recognize regions from mutagenesis in the coat protein (CP) gene are important for particle assembly and the expression of viral in CP expression systems like bacteria such as E.coli or yeast as well as their assembly into virus-like particles (VLP). In this paper, we report the expression and assembly of VLP in cells of E. coli expressing the CMVCP gene. 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. 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. The results showed that the CP gene was expressed in the prokaryotic system successfully and was assembled into the CMV-like particle.
