Construction and identification of the eukaryotic expression vector of vaccinia virus expressing human interleukin-2

Abstract

OBJECTIVE: Cancer gene therapy using interleukin-2 (IL-2) has generated much interest because of the potent antitumor effect of this cytokine. There is ongoing research into one promising new gene therapy approach for cancer using the vaccinia virus vector. The purpose of this study was to construct a vaccinia eukaryotic expression vector, pMJ601, which contains human IL-2 (hIL-2; pMJ601hIL-2) and can be used for the treatment of gastric carcinomas. METHODS: Genetic engineering techniques such as plasmid extraction, agarose gel electrophoresis, restriction analysis, ligation, preparation of competent cells, transformation and DNA sequence analysis were used to clone the hIL-2 gene into pBluescript II SK+/– and pMJ601 and identify these vectors. RESULTS: Fragments of hIL-2 DNA from pLXSN from an EcoR1–BamHI restriction enzyme digest were successfully cloned into pBluescript II SK+/–. In addition, hIL-2 DNA from pBluescript II SK+/– with a SalI-BamHI restriction enzyme digest was also successfully cloned into pMJ601. CONCLUSION: Constructing a pMJ601 vector that expresses the hIL-2 gene is an important step toward being able to treat gastric carcinoma using gene therapy.