PLASMID FOR EXPRESSION OF APOPTIN IN Escherichia coli

The chicken anemia virus (CAV) Apoptin can induce apoptosis specifically in tumor cells and not in normal cells. Therefore, this shows a promising use for cancer gene therapy in the future. However, the mechanism of apoptin-induced apoptosis in tumor cells has not been well characterized. We tried to construct p-Ec-Apo (Apoptin expression plasmid for Escherichia coli) for further use, especially to explore the molecular aspect and therapeutically strategy of Apoptin. Aim of this study is to construct plasmid for expression of Apoptin in Escherichia coli. A complete open reading frame of the Apoptin gene of CAV/wild type was amplified by PCR. The PCR product was then purified from the agarose using QIAquick Gel Extraction Kit (Qiagen), subcloned into the respective sites of pETBlue-1 (Novagen). The molecular clone was transformed into Escherichia coli, purified, and sequenced. The nucleotide sequence was analyzed by CLC Main Workbench (CLC Bio). Results: The clone thus obtained, p-Ec-Apo, was successfully constructed. Keywords : Apoptin, Escherichia coli