Identification of the structural proteins of VP1 and VP2.pdf

PI d R X S A R R A A K M S M 1 i C 1 2 v 2 e t o T a a s a d n n 0 dJournal of Virological Methods 171 (2011) 323–328 Contents lists available at ScienceDirect Journal of Virological Methods journa l homepage: www.e lsev ier .com/ locate / jv i romet rotocols dentification of the structural proteins of VP1 and VP2 of a novel mud crab icistrovirus ui Zhang1, Jianguo He1, Hongjun Su, Chuanfu Dong, Zhixun Guo, Yujie Ou, iexiong Deng, Shaoping Weng ? tate Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-Sen (Zhongshan) University, 135 Xingang Road West, Guangzhou 510275, China rticle history: eceived 25 February 2010 eceived in revised form 24 August 2010 ccepted 8 September 2010 vailable online 17 September 2010 a b s t r a c t Mud crab dicistrovirus (MCDV), a newly identified single-stranded positive RNA virus, is an important pathogen that causes serious economic losses to mud crab aquaculture. In this study, MCDV was puri- fied, and three structural proteins of MCDVwere separated by SDS. The N-terminal 15 amino acids were sequenced and alignedwith themain structural proteins of other dicistrovirus. The three structural proteins were named VP1, VP2 and VP3. Monoclonal antibodies (MAbs) against the two main structuraleywords: ud crab dicistrovirus (MCDV) tructural protein onoclonal antibodies (MAbs) proteins, VP1 and VP2, were prepared, and the two structural proteins were then identified using these MAbs. The results of Western blot analyses demonstrated that five MAbs recognised VP1 and two recog- nised VP2. The results of immunogold transmission electronmicroscopy (IEM) revealed that the epitopes of the two structural proteins recognised by the MAbs were located at the outer surface of the virions, which suggested that the two structural proteins areMCDV capsid proteins. The identification of the two structural proteins of MCDV is useful for studying their functions, as well as the mechanism of infection CDVand the pathogenesis of M . Introduction Th