Available online www.ijpras.com International Journal of Pharmaceutical Research & Allied Sciences, 2016, 5(3):343-349 Research Article ISSN : 2277-3657 CODEN(USA) : IJPRPM 343 Generation of mini-Tn7 transposone by cloning of CPV-VP2 into eukaryotic expression cassette, in E. coli Mohammad Sadegh Hashemzadeh 1 , Seyed Jafar Mousavy *2 , Ruhollah Dorostkar 3 , Fatemeh Fotouhi 4 and Firouz Ebrahimi 5 1 Ph.D Candidate in Nanobiotechnology, Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran 2 Assistant Professor of Biochemistry (Ph.D), Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran 3 Assistant Professor of Virology (Ph.D), Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran 4 Associate Professor of Virology (Ph.D), Department of Influenza and other Respiratory viruses, Pasteur Institute of Iran, Tehran, Iran 5 Assistant Professor of Clinical Biochemistry (Ph.D), Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran * Corresponding E-mail: jmosavi@ihu.ac.ir _____________________________________________________________________________________________ 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. Keywords: Canine parvovirus, VP2, expression cassette, pFastBac1 _____________________________________________________________________________________________ INTRODUCTION Canine parvovirus (CPV) is a member of the Parvovirus genus in the family of Parvoviridae. This family are spherical, nonenveloped, T=1 icosahedral viruses that infect a wide range of natural hosts. CPV first appeared in the late 1970s and is a natural pathogen of dogs [1,2,3] and encapsidates a single-stranded DNA genome of approximately 5 kb. CPV particles have a diameter of 25 nm and are composed of three proteins, VP1, VP2, and VP3 [4]. VP2 is the major component of the viral capsid and contains 584 amino acid residues. About 90% of the