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 adult kidney and occurs in its sporadic and hereditary form. Approximately 57% of all renal carcinomas of the clear cell type analyzed revealed a mutation in the von Hippel-Lindau disease (VHL) gene. In the present work, temperature gradient gel electrophoresis (TGGE) is presented as a rapid and precise polymerase chain reaction (PCR)-employing methodology 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 known mutations in the VHL gene are detectable. This calculated figure appears to be in agreement with the experimental results. Primary difficulties in analyzing exon 1 of the VHL gene were overcome by the employment of psoralen-cross-linked PCR fragments. In addition, TGGE analysis was used in screening for possible mutations in thirteen renal carcinoma samples. With this protocol TGGE is successfully added to the array of methods for the screening of VHL mutations.
Wenzel
M
M
Knochenmarkspenderzentrale, Heinrich-Heine-Universität Düsseldorf, Germany.
Enczmann
J
J
Uhrberg
M
M
Hernández
A
A
Wiese
U
U
Ackermann
R
R
Schmitz-Draeger
B
B
Ebert
T
T
Wernet
P
P
D000368:Aged; D002292:Carcinoma, Renal Cell / Q000235:genetics; D004252:DNA Mutational Analysis; D004586:Electrophoresis / Q000379:methods*; D005260:Female; D016147:Genes, Tumor Suppressor; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction; D013696:Temperature; D006623:von Hippel-Lindau Disease / Q000235:genetics*