Molecular cloning of an enhancer binding protein: isolation by screening of an expression library with a recognition site DNA

Cell. 1988 Feb 12;52(3):415-23. doi: 10.1016/s0092-8674(88)80034-5.


A novel strategy has been used to isolate a cDNA clone that encodes a DNA binding domain whose recognition properties overlap those of the mammalian transcription factors H2TF1 and NF-kappa B. These two factors are distinguished by their cell type distributions and their relative affinities for related sequence elements in the enhancers of the major histocompatibility complex (MHC) class I and immunoglobulin kappa chain genes. The human cDNA clone was detected by screening a lambda phage expression library with a binding site probe derived from the MHC enhancer. The phage encoded fusion protein binds specifically to both the MHC and kappa gene enhancers. The cDNA hybridizes to a single copy gene that is expressed as a 10 kb mRNA in both B and non-B cells. The strategy used in this study may prove generally useful in the cloning and analysis of sequence-specific DNA binding proteins.

Publication types

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

MeSH terms

  • Animals
  • Bacteriophage lambda / genetics
  • Base Sequence
  • Binding Sites
  • Cloning, Molecular
  • DNA
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Enhancer Elements, Genetic*
  • Genes, MHC Class I*
  • Mice
  • Nucleic Acid Hybridization
  • Recombination, Genetic
  • Transcription Factors / metabolism


  • DNA-Binding Proteins
  • Transcription Factors
  • DNA