Genome Sequences Of Basella Rugose Mosaic Virus Isolated From Mirabilis Jalapa And The Construction Of Prokaryotic Expression Vector With Antibody Preparation Of CP Gene

Biological characteristics of Basella rugose mosaic virus(BaRMV), which infected Mirabilis jalapa, were systematically studied, and partial genome was sequenced. We constructed prokaryotic expression vector and prepared antibody of CP gene of BaRMV. The results showed that:1、Mirabilis jalapa infected showed systemic infection syptoms, such as deformed and mosaic leaf; The EM observation showed that the virus particles was13x700～800nm in length; Inoculation experiment showed that Nicotiana benthamiana were successfully inoculated. Compared to the mechanical inoculation method, the injection inoculation method showed a higher efficient (0%and76%in infection ratio, respectively). The ELISA and RT-PCR were detected for inoculated plants showed that the incidence reached more than76%.2、The genomic sequence (except5’-UTR and patial of Pl ene) of BaRMV-MJ were determinated, Sequence analyzing showed that there were great differences existed in the cleavage sites of the BaRMV-MJ and other isolates of CI,6k2, NIa-Pro and Nib protein. The motif of DAG’, which was involved in aphid transmission related, was turned into’NAG’. However, the "pipo" motif which presented as GGAAAAAA(A) was found in P3region. Homology analysis and phylogenetic tree analysis of the patial gene of BaRMV-MJ, such as CP, CI gene based on nucleotide and amino acid showed that the BaRMV-MJ should be one isolate of BaRMV.3、Prokaryotic expression vector of pET-30a-CP was constructed successfully according to RT-PCR and cloning. Polyclonal antibody was prepared by immuning the new zeland white rabbit with CP protein. The antibody was tested by using ID-Elisa and western blot metord. Results showed that the antibody had a proper titer and specificity which could be used for the subsequent testing and provide reliable test method for related researches of BaRMV in Yunnan province.