Analysis of genetic mutations in human lactate dehydrogenase-A(M) deficiency using DNA conformation polymorphism in combination with polyacrylamide gradient gel and silver staining

Biochem Biophys Res Commun. 1991 Oct 31;180(2):1083-90. doi: 10.1016/s0006-291x(05)81177-5.

Abstract

Human lactate dehydrogenase (LDH)-A mutant gene was analyzed by polymerase chain reaction - DNA conformation polymorphism (DCP). We used polyacrylamide gradient gel and silver staining procedures for DCP analysis and observed abnormal migration patterns in individuals heterozygous for LDH-A deficiency. Further sequence determination of the mutant alleles consistently resulted in detection of base substitutions, a G to T transversion at codon 328 (GAG----TAG), and synonymous substitutions at codon 115, 160 and 172. Such mutations were easily detectable using the DCP technique. The DCP technique using the polyacrylamide gradient gel and silver staining method seems likely to be useful for the rapid screening of mutations and for further genotype detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • DNA / chemistry
  • DNA / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Exons
  • Genetic Carrier Screening
  • Humans
  • Introns
  • Isoenzymes
  • L-Lactate Dehydrogenase / deficiency
  • L-Lactate Dehydrogenase / genetics*
  • Molecular Sequence Data
  • Mutation*
  • Nucleic Acid Conformation
  • Oligodeoxyribonucleotides
  • Polymerase Chain Reaction / methods
  • Polymorphism, Genetic

Substances

  • Isoenzymes
  • Oligodeoxyribonucleotides
  • DNA
  • L-Lactate Dehydrogenase