Maurya & Rai (2017) Biotechnology International 10(2): 52-58 52 ©Biotechnology Society www.bti.org.in ISSN 0974-1453 Research article CHICKEN ANEMIA VIRUS VP3 GENE CLONED IN pVAX1 VECTOR CAUSES APOPTIC ACTIVITY IN HeLa CELLS Raushi Maurya 1 and Anant Rai 2* 1 Department of Biotechnology, Mewar University, Chittorgarh, Rajasthan. 2 Institute of Biotechnology & IT, 197-198, Mudiya Ahmadnagar, Bareilly-2453122, UP. *Corresponding author: raia48@gmail.com ABSTRACT VP3 protein gene of chicken anemia virus cloned in pVAX1 was analysed for its apoptotic activity using HeLa cells in culture. It was found to produce high level of apoptosis when analysed by DNA fragmentation assay, acridine orange-ethidium bromide staining, caspase assay, annexin V binding assay, Tunel assay and MTT cytotoxic assay. Keywords: VP3, apoptin, pVAX1, caspase, annexin V binding. INTRODUCTION The 2.3 kb chicken anemia virus (CAV) genome encodes three viral proteins VP1, VP2, VP3. VP1 is major capsid protein and VP2 is probably a non- structural protein found in the cells in the early stage of virus replication cycle (Noteborn et al., 1995). VP1 and VP2 are protective proteins that induce neutralising antibodies (Kock et al., 1995). The third viral protein VP3 is a 13 kDa protein that shows apoptic activity and is able to induce apoptosis within infected chicken cells and human tumor cell lines (Noteborn et al., 2004). VP3 has been shown to not only induce apoptosis when introduced into the precursors of chicken thymocytes, but has been found to specifically kill human cancer cells and transformed cells without affecting the proliferation of normal cells (Lee et al., 2012). Apoptin causes apoptosis in various human tumors and transformed cells (Danen Van Oorschot et al., 1997). Apoptin is a promising and ideal agent for cancer gene therapy owing to its intrinsic specificity and the inherent low toxicity (Gueden et al., 2004). Because of its death inducing abilities, the VP3 gene product was renamed apoptin. Interestingly, the apoptic activity of apoptin in not restricted to chicken thymocytes (Danen Van Oorschot et al., 1997). Unlike other tumor suppressor or proapoptic genes, theapoptin does not rely on the function of p53, BCL- 2, Bax, FADD, or Caspase-8 (Tavassoli et al., 2005; Liu et al., 2006). Thus apoptin may be used to treat a broad spectrum of tumors that differ in mutational status of p53 and the levels of expressions of apoptotic regulators. The ability of apoptin to induce apoptosis has been demonstrated in more than 70 human cancer cell lines (Backendorf et al., 2008; Noteborn et al.,