Analysis of the p53 tumor suppressor gene by pyrosequencing.
Tumor suppressor genes are implicated in cell cycle progression. Inactivation of these genes predominantly occurs through mutations and/or allelic loss that involves both alleles. With inactivation by multiple mutations in a single gene, cloning of the amplified gene is necessary to determine whether the mutations reside on one or both alleles. Using pyrosequencing, a recently developed approach based on sequencing-by-synthesis, we studied genetic variability in the p53 tumor suppressor gene and could quantify the ratio between the mutated and wild-type amplified fragments. Furthermore, this sequencing technique also allows allelic determination of adjacent mutations with no cloning of amplified fragments.
Ahmadian
A
A
Royal Institute of Technology, Stockholm, Sweden.
Lundeberg
J
J
Nyrén
P
P
Uhlén
M
M
Ronaghi
M
M
D000483:Alleles; D004247:DNA / Q000096:biosynthesis; D017931:DNA Primers; D003854:Deoxyribonucleotides / Q000378:metabolism; D049454:Firefly Luciferin / Q000378:metabolism; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods; D012878:Skin Neoplasms / Q000235:genetics; D013430:Sulfate Adenylyltransferase / Q000378:metabolism
D017931:DNA Primers; D003854:Deoxyribonucleotides; D049454:Firefly Luciferin; D004247:DNA; D008156:Luciferases; D013430:Sulfate Adenylyltransferase