Molecular cloning of an enhancer binding protein: isolation by screening of an expression library with a recognition site DNA.
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.
Singh
H
H
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
LeBowitz
J H
JH
Baldwin
A S
AS
Sharp
P A
PA
D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005805:Genes, MHC Class I; D051379:Mice; D009693:Nucleic Acid Hybridization; D011995:Recombination, Genetic; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; D014157:Transcription Factors; D004247:DNA