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A five‐basepair deletion (7118 delTTTTA) identified within neurofibromatosis NF1 exon 39
Author(s) -
Rodenhiser David I.,
Jung Jack H.,
Gillet Jane M. R.,
Hovland Ken,
Andrews Joseph,
Ainsworth Peter J.,
CoulterMackie Marion,
Singh Shiva M.
Publication year - 1997
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1997)9:5<473::aid-humu15>3.0.co;2-#
Subject(s) - heteroduplex , biology , genetics , exon , microbiology and biotechnology , restriction enzyme , frameshift mutation , polymerase chain reaction , mutation , complementary dna , stop codon , sequence analysis , base pair , dna , gene
We have screened a panel of neurofibromatosis type 1 (NF1) patients representing 85 kindred from Ontario, Canada. DNA was extracted and polymerase chain reaction (PCR) used to amplify a 285‐bp DNA fragment containing 39 (Abernathy et al., 1994). Heteroduplex analysis (Ainsworth et al., 1994) of the PCR products revealed the presence of a heteroduplex variant from one patient (#70‐630). PCR product sequenced directly and also cloned revealed a five‐base deletion (7118 delTTTA). Computer analysis of the resulting amino acid sequence showed that this deletion alters the reading frame of exons 39 and 40, where a novel stop codon is generated at position 7215 of the cDNA sequence. We also used restriction enzyme analysis to verify the location of this mutation since the deletion eliminates a Dra I site (TTT/AAA) partially contained within this sequence. Dra I digestion of this PCR product gave 168‐ and 117‐bp fragments from the normal allele but Dra I did not digest product amplified from the mutation allele. DNA from the patient's affected mother also diplayed the heteroduplex variant and loss of this Dra I site.