CLONING AND RECOMBINANT EXPRESSION OF MOUSE COAGULATION-FACTOR-X

Engineering of recombinant coagulation factor X variants, which can be activated by tumor-associated proteinases may lead to the development of new therapeutic molecules. However, the evaluation of such variant s requires an appropriate animal model. Therefore, we isolated the com plete coding sequence of mouse coagulation factor X from mouse liver c DNA by polymerase chain reaction. The deduced amino acid sequence code s for a prepro protein of 481 amino acids homologous to factor X seque nces from various species. Recombinant mouse factor X was expressed in human embryonic kidney cells and secreted into cell culture supernata nt as zymogen, which could be converted to catalytically active factor Xa by Russell's viper venom. Purified recombinant mouse factor X rest ored coagulation in human factor X deficient plasma, demonstrating tha t mouse factor X is able to functionally interact with the human blood coagulation system. Recombinant mouse factor X opens the possibility to analyze therapeutically useful variants in the mouse system. (C) 19 98 Elsevier Science Ltd.