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CHARACTERIZATION OF THE MUTATION CONCERNING THE GENES WHICH CODIFY THE COAGULATION FACTORS
 
 

Together with the diagnosis, the mutation's identification and the study of their functional effects are also important because they represent a model of what is present in nature and could offer information on the structure-function relation in coagulation factors.

These studies are developed through:
1) The study of the structure of the mutant protein's crystal:
  - Localization of the variation found in the three-dimensional structure of the protein in order to assess the importance of the structural level and its influence on the functionality of the protein
 

- The coordinates of the protein's crystals are extracted from a Protein Data Bank. The structure of the protein is visualized through the software P INSIGHT II 98.0.
2) In-vitro studies:
  - Production of normal and mutant constructions through the Site-Directed Mutagenesis technology.
  - Production of the recombinant protein through transitory or stable transfection of said constructs in appropriate cell lines through the method of the electroporation.
  - Intra- and extra-cellular kinetic analysis of the recombinant protein that has been mutated with the procedure of immunoprecipitation and pulse-chase labeling using S35-marked methionine in order to check a possible deficiency in the secretion of the protein.
  - Immunofluorence with specific antibodies.
  - Cleansing of the mutated recombinant protein through FPLC or HPLC and further biochemical characterization of said protein.
   
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