Use of gradient denaturing gels to determine mutational spectrum in human cells.
Based on the fact that mutagens induce specific patterns of gene mutations, this paper outlines a method to allow discrimination among mutagen-treated populations. The technique should allow direct screening of human tissue for genetic change, using human peripheral blood lymphocytes deficient in the enzyme hypoxanthine guanine phosphoribosyl transferase. The method is based on gradient denaturing gel electrophoresis, which separates short DNA molecules according to their melting properties. The melting behavior of DNA fragments is extremely sequence-dependent, and DNAs with single basepair substitutions often migrate differently. Even DNA fragments with the same basepair substitutions at different locations in the molecule have been resolved. Gradient-denaturing gel electrophoresis has the capacity to separate mutant DNA on the basis of the nature and position of the mutation.
D001483:Base Sequence; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D009154:Mutation; D009691:Nucleic Acid Denaturation
D004269:DNA, Bacterial; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase