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Screening for renal carcinoma associated mutations in the von Hippel‐Lindau tumor suppressor gene by temperature gradient gel electrophoresis
Author(s) -
Wenzel Michael,
Enczmann Jürgen,
Uhrberg Markus,
Hernández Ana,
Wiese Ulrich,
Ackermann Rolf,
SchmitzDraeger Bernd,
Ebert Thomas,
Wernet Peter
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150180109
Subject(s) - temperature gradient gel electrophoresis , renal cell carcinoma , microbiology and biotechnology , von hippel–lindau disease , gene , biology , exon , mutation , tumor suppressor gene , cancer research , carcinoma , polymerase chain reaction , gel electrophoresis , gene mutation , genetics , pathology , carcinogenesis , disease , medicine , 16s ribosomal rna
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.