SCREENING FOR RENAL-CARCINOMA ASSOCIATED MUTATIONS IN THE VON-HIPPEL-LINDAU TUMOR-SUPPRESSOR GENE BY TEMPERATURE-GRADIENT GEL-ELECTROPHORESIS

Renal cell carcinoma is the most common neoplastic disease of the adul t kidney and occurs in its sporadic and hereditary form. Approximately 57% of all renal carcinomas of the clear cell type analysed revealed a mutation in the von Hippel-Lindau disease (VI-IL) gene. In the prese nt work, temperature gradient gel electrophoresis (TGGE) is presented as a rapid and precise polymerase chain reaction (PCR)-employing metho dology for the detection of mutations in the VHL gene. The theoretical efficiency of TGGE to detect mutations in every base pair of the gene was calculated. According to computer analysis, at least 92% of all k nown mutations in the VHL gene are detectable. This calculated figure appears to be in agreement with the experimental results. Primary diff iculties in analysing exon 1 of the VHL gene were overcome by the empl oyment of psoralen-cross-linked PCR fragments. In addition, TGGE analy sis was used in screening for possible mutations in thirteen renal car cinoma samples. With this protocol TGGE is successfully added to the a rray of methods for the screening of VHL mutations.