Clonagem e sequenciamento das regiões codificantes das proteínas large e small do capsídeo do vírus do mosaico severo do caupi (CPSMV) para a produção de viral like particles

Abstract

The present work reports the sequencing of the CPSMV RNA-2 polyprotein, having seen a constant search for biotechnological solutions to supply the demands that fit or depend on the development of approaches involving the biology of molecules, such as DNA sequencing, VLPs and vaccines. In this aspect, a sequencing of an unknown region was performed in some strains already published in its RAN-2, with some Nordestins, from CPMSV, where there is only a partial sequence of the polyprotein coding region resulting from this RNA-2 fragmentation of this virus, this one by its turn is the one that, after post-translational modifications, gives rise to a cell-to-cell movement protein and two protein subunits, a larger (~41 kDa)(CPL) and a smaller (~21 kDa)(CPS). With the advent of VLP technology, the search for proteins that fit this approach has been growing, given the wide applicability of this technology. Protein subunits (CPL) and (CPS) present this possibility of formation, since close phylogenetic viruses such as CPMV already have until the commercialization of the VLP formed by it. With this premise and the identification of databases such as the Genbank (NCBI), it was possible to analyze the regions that are unknown in some strains and compare them with others that already have partial known sequences of the region that originate the proteins (CPL) and (CPS), through multiple alignments, using software such as Blastn and Clustalw. And through molecular biology techniques, CR and molecular cloning in the pGEM-T cloning vector, it was possible to amplify the unknown regions present in the partial sequences already published.