The Experimental Research On The Therapy Of Pancreatic Cancer By TK Gene That Activated By Egr-1 Promoter

1. To construct AD vector containing TK gene controlled by Egr-1promoterObjectives To construct recombinant adenovirus vector pAdEgr-1-TK of the suicide gene that containing herpes simplex virus thymidine kinase gene(HSK-TK or TK),which is controlled by the early growth response-1(Egr-l).The vector was marked by green fluorescent protein(GFP),so that the transfection rate and growth of virus could be recorded by observing GFP in the fluorescence microscope. Methods Egr-1 was amplificated according to the template plasmid vector PCl-neo-Egr-1-TK(pET), then digest by XbaI in the upstream and XhoI in the downstream.The 445bp Egr-1 fragment was obtained after separated the product in the 1.0% agarose gel.Then recombinant plasmid pAdTrack/Egr-1 was obtained after cloning the gene into the shuttle plasmid pAdTrack.pET turned to TKcDNA after digest and complemented by EcoRI,and digest again by Not I.then together with pAdTrack/Egr-1, digest by EcoRI and Not I, the two product were ligated and recombinant plasmid pAdTrack/Egr-1-TK was created.In the bacteria...