Study On Recombinant Nonglycosylated Urokinase-type Plasminogen Activator

Part 1 Construction of single-chain urokinase-type plasminogen activator(scu-PA) mutant geneObjective To construct nonglycosylated, thrombin resistant, high molecular weight pro-urokinase mutant.Method With PCR amplification, glycosylation site was deleted by Asn302-Ala302 mutagenesis. Thrombin cleavage site was eliminated by Argl56-Lysl56 mutagenesis. Results DNA molecular weight of PCR product was 1296 daltons as determined by agarose gel electrophoresis. The mutant and puc19 cloning vector were pretreated by HindIII, XbaI digestion. After ligation, they were transformed into DH5α competent cells (Escherichia coli.). PCR products cloned into puc19 vectors can be...