Cloning of the Ssap-NtrB prokaryotic gene into the eukaryotic expression vector pcDNA3.1 / V5 / His B vector

Nelin Hacıoğlu, Feray Köçkar


Suicide gene therapy has recently emerged as a method used in cancer treatments.  These therapies utilized enzymes that are expressed in the cell.  In this study, Staphylococcus saprophyticus supsp. saprophyticus Nitroreductase gene (Ssap-NtrB) was subcloned into the eukaryotic expression vector namely pcDNA3.1 / V5 / His B. For this purpose, Nitroreductase gene region was firstly amplified from the pET14B vector using PCR strategy and cloned into the pGEM-T-Easy vector.  After this step, the Ssap-NtrB gene was restricted with KpnI/ApaI and was ligated into pcDNA3.1 / V5 / His B vector.  Recombinant colonies were verified using KpnI/ApaI restriction enzymes. As a result, the Ssap-NtrB gene was cloned into pcDNA3.1/V5/His B vector and was readyfor use in suicide gene therapy in eukaryotic human cancer cells.

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