"Hot spots" mutation analysis of p53 gene in gastrointestinal cancers by amplification of naturally occurring and artificially created restriction sites

Pao Huei Chen, Shyr Yi Lin, Chung Kwe Wang, Yi Jen Chen, Te Chuan Chen, Jan Gowth Chang

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We developed a rapid, simple method to detect "hot spot" point mutations of the p53 gene. A DNA fragment from cancer tissue of a surgical specimen was selectively amplified with specific oligonucleotide primers and then digested with restriction enzymes that recognized artificial or naturally occurring restriction sites. To detect mutations in codons 273 and 245, we created artificial Bst U1 and Bgl I sites by introducing a single nucleotide mismatch into the respective mutagenesis primers. We used the naturally occurring restriction sites of Msp I, Hae III, and Hha I to detect mutations in codons 248, 249, and 175, respectively. In 74 cases of gastrointestinal cancer, 5 of 35 colorectal cancers showed mutations; 1 of 15 cases of gastric cancers showed mutation; and 1 of 24 hepatocellular carcinomas showed mutation. This nonradioactive method is an accurate and clinically useful way to detect hot-spot point mutations of the p53 gene in gastrointestinal cancers.

Original languageEnglish
Pages (from-to)2186-2191
Number of pages6
JournalClinical Chemistry
Volume39
Issue number10
Publication statusPublished - 1993
Externally publishedYes

Keywords

  • DNA probes
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Clinical Biochemistry

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