Construction And Evaluation Of Targeted Plasmid Vector PcDNA3.1(-)Cp-CD-TK

ObjectiveTo Construct targeted plasmid vector pcDNA3.1(-)Cp-CD-TK which contains CEA promoter. To study CD-TK expression driven by CEA specific promoter and its lethal effect in CEA-positive colorectal carcinoma cell.MethodsWe Constructed the targeted plasmid vector pcDNA3.1(-)Cp-CD-TK which contains CEA promoter by PCR, enzyme restriction, ligation and other molecular biology method. We then transfected the plamid into the CEA-positive cells (SW480) and CEA-negative cells (Hela) respectively. Expression of CD-TK gene was detected by RT-PCR. MTT assay was used to detect the CEA-specific lethal effect of the Cp-CD-TK system.ResultsThe targeted plasmid vector pcDNA3.1(-)Cp-CD-TK was constructed successfully. The CEA-positive cells (SW480) showed CD-TK expression and were sensitive to prodrug after transfection. The CEA-negative cells (Hela) showed negative result.ConclusionsCD-TK expression of pcDNA3.1(-)Cp-CD-TK was CEA-specific and this targeted pladmid vector cast new light on colorectal carcinoma gene therapy.