Restriction isotyping of human apolipoprotein E by gene amplification and cleavage with HhaI

J Lipid Res. 1990 Mar;31(3):545-8.


We have used restriction isotyping (restriction enzyme isoform genotyping) for rapid typing of common apolipoprotein E isoforms (E2, E3, E4). ApoE restriction isotyping used oligonucleotides to amplify apolipoprotein E gene sequences containing amino acid positions 112 and 158. The amplification products were digested with HhaI and subjected to electrophoresis on polyacrylamide gels. Each of the isoforms was distinguished by a unique combination of HhaI fragment sizes that enabled unambiguous typing of all homozygotic and heterozygotic combinations. HhaI cleaves at GCGC encoding 112arg (E4) and 158arg (E3, E4), but does not cut at GTGC encoding 112cys (E2, E3) and 158cys (E2).

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Apolipoproteins E / genetics*
  • Apolipoproteins E / isolation & purification
  • Base Sequence
  • DNA / isolation & purification
  • DNA / metabolism
  • Deoxyribonucleases, Type II Site-Specific / metabolism*
  • Gene Amplification*
  • Humans
  • Leukocytes / metabolism
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Polymorphism, Genetic


  • Apolipoproteins E
  • DNA
  • Deoxyribonucleases, Type II Site-Specific
  • GCGC-specific type II deoxyribonucleases