Abstract

The VP2 protein of canine parvovirus (CPV) is the main part of capsid and attachment ligands for entry intospecific and cancerous cells through transferrin receptors (TfRs). Expression of VP2 alone results in assembly of a typically-sized virus like particle (VLP) in insect cells for therapeutic purposes. Towards this goal, the first step is to construct an expression cassette in pFastBac1 donor vector for transposition of VP2 into bacmid shuttle vector using Bac-to-Bac baculoviral system. So, in this research, we generated new recombinant pVP2FastBac1 enablingsite-specific transposition of VP2 into bacmid. The full-length of CPV-VP2 gene (1755 bp) was isolated by PCR amplification using specific primers and cloned firstly into RBC T/A cloning vector and then subcloned into the corresponding restriction sites of pFastBac1 donor plasmid vector. Then the accuracy of cloning process in these vectors was evaluated by PCR and enzymatic digestion analysis. Successful cloning of CPV-VP2 gene into eukaryotic expression cassette of pFastBac1 donor vector was confirmed by PCR and enzymatic digestion. In other words, mini-Tn7 transposone was generated successfully. In this study, the mini-Tn7 transposone containing CPV-VP2 gene was constructed. It is required for transposition of the interest gene into bacmid DNA in order to express it in insect cell.