Abstract:
Until now, the study of the multiple endocrine
neoplasia type 1 (MEN1) gene in patients suspected of
having the disease was expensive and laborious due to the
large size of the gene. We have optimized the conformation sensitive gel electrophoresis (CSGE) technique to analyze
by four rather simple multiplex PCR reactions, and a single
electrophoresis run, the entire coding region of the MEN1
gene, plus the exon¿intron boundaries. This improvement
of the CSGE technique was confirmed as an effective proce dure for screening for the MEN1 gene by detecting ten
previously known MEN1 gene mutations and four polymor phisms. The MEN1 gene of 12 patients with unknown mu tations was then screened, and an abnormal CSGE profile
was identified in 10/12 cases. Subsequent DNA sequencing
demonstrated 3 of them to be novel mutations (E45K,
4479delACAG, 6073insC) and 7 to have been previously
reported; in the remaining 2 patients, we confirmed the
absence of any alteration of the coding sequence of MEN1.
Mutation screening of the MEN1 gene using CSGE was
demonstrated to be a fast, simple, and inexpensive method
to study patients suspected of having MEN1 disease.
