[Experimental studies on somatic gene therapy for diabetes. I. Structuring of recombinant from human insulin gene and...

To develop a model somatic gene therapy system for diabetes, we constructed a human insulin expression vector in non B cells. As the first step, an insulin cDNA fragment of 260 bp, generated by a complet digestion of PBCA with EcoR I and BamH I, was inserted into the EcoR I/BamH I site of plasmid PBS.SK by ligation of cohevise-ended DNA to construct transition plasmid PBS.INS. Then the plasmid PBS.INS was completly digested by Hind III and Xbal I. The small DNA fragment containing insulin cDNA gene was subcloned to the expression plasmid PRC/CMV to form recombinant PRC/CMV.INS.