Human prothrombin kringle with endothelial cell growth inhibitory activity vector and host thereof

摘要

PURPOSE: Provided are a human prothrombin kringle protein with endothelial cell growth inhibitory activity, a recombinant vector for its expression and a transformed E. coli by the vector. The protein is used for the treatment of diseases involving the abnormal growth of endothelial cell such as cancer and arthritis. CONSTITUTION: The preparation method comprises the steps of: amplifying cDNA fragments of each human prothrombin kringle 1 and 1-2 by a polymerase chain reaction with cDNA of human prothrombin as a template and using the oligo-nucleotide primer synthesized based on the amino acid sequences of N-and C-terminals of each human prothrombin kringle 1 and 1-2; making pEF-32hK1 and pEF-hK1-2 vector by cloning the above amplified cDNA fragments to the E.coli expression vector; making each E.coliE transformant (KCCM-1082) and E.coli transformant (KCCM-10183) transformed woth vector pEF-32K1 and pEF-hK1-2 respectively and separating and purifying tagged human prothrombin kringle 1(32bK1) through ion-exchange chromatography of the excessively expressed E.coli extracts and using histidine tag; and treating the above human prothrombin kringle 1 with thrombin and applying affinity chromatography to obtain pure human prothrombin kringle 1.