B-raf polynucleotides
The invention relates to mutations in B-Raf gene products. The mutations described are identified in human tumors of natural origin. These mutations are associated with cancerous phenotypes and can be used as a basis for the diagnosis of cancer, cancerous cells or a predisposition to cancer in human subjects, and the development of anti-cancer therapeutics.
1. An isolated nucleic acid that hybridizes under stringent hybridization conditions to SEQ ID NO: 2,
wherein the nucleic acid is the complement of a polynucleotide that encodes a variant B-Raf polypeptide having kinase activity and comprising Glu or Asp at the position corresponding to position 599 of SEQ ID NO: 1,
wherein said stringent hybridization conditions comprise an overnight incubation at 42° C. in a solution comprising 50% formamide, 5X SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5X Denhardt's solution, 10% dextran sulphate, and 20 pg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1X SSC at 65° C.
2. An isolated nucleic acid which comprises a polynucleotide encoding a variant B-Raf polypeptide having at least 95% identity to SEQ ID NO: 1, wherein the polypeptide comprises a Glu or Asp at the position corresponding to position 599 of SEQ ID NO: 1 and has kinase activity.
3. The complement of the isolated nucleic acid of claim 2 .