Mutation analysis of the NF1 gene

摘要

A method for mutation analysis of the neurofibromatosis 1 (NF1) gene of a patient includes extracting DNA from peripheral blood lymphocytes of the patient, establishing an EBV transformed B-lymphoblastoid cell line using lymphocytes from the patient, treating the EBV transformed B-lymphoblastoid cell line culture with puromycin, extracting RNA from cultures of the cell line immediately, amplifying the RNA using suitable primers, and obtaining peptide fragments by means of in vitro transcription/translation of the amplified fragments. The invention also relates to the identification of new hotspots and specific NF1 mutations. The invention also includes diagnostic kits for the detection of described specific mutations and hotspot domains, compounds correcting the structure of specific mutated NF1 proteins and in vitro and in vivo systems that may be used to screen for these therapeutic compounds.