Produzione di fattore di Hageman (FXII umano) in cellule di scimmia mediante espressione transiente del suo cDNA

- Human coagulation factor XII (FXII) is a serine protease involved in a wide variety of functions, including blood coagulation, fibrinolysis, bradykinin production and complement activation. FXII consists of several autonomous structural regions similar to those present in other serine proteases. With the intent of assessing the contribution of each structural domain to the biological properties of the enzyme, we have built an experimental model that allows the expression of FXII cDNA in eukaryotic cells. The fulll-ength FXII cDNA was constructed and cloned into the shuttle vector pSVL under the SV40 promoter control. Simian cells (COS) were transfected with either the recombinant or the wild-type vector. At different times after transfection the cells and the conditioned medium were harvested and tested for the production and secretion of the recombinant FXII. FXII cDNA transcription and translation were demonstrated by Northern transfer, indirect immunofluorescence and Western blot, while the functional activity of the recombinant protein was proved by a coagulation assay. © 1990 Accademia nazionale dei Lincei.