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. 1992 May 11;20(9):2257–2263. doi: 10.1093/nar/20.9.2257

Determination of the DNA sequence recognized by the bHLH-zip domain of the N-Myc protein.

R Alex 1, O Sözeri 1, S Meyer 1, R Dildrop 1
PMCID: PMC312339  PMID: 1594445

Abstract

The DNA-binding domain of the murine N-Myc protein, comprising the basic helix-loop-helix-zipper (bHLH-zip) region was expressed as a fusion protein in E. coli. The affinity purified glutathione-S-transferase-N-Myc fusion protein (GST-N-MYC) was used to select the N-Myc specific DNA-recognition motif from a pool of random-sequence oligonucleotides. After seven rounds of binding-site selection, specifically enriched oligonucleotides were cloned and sequenced. Of 31 individual oligonucleotides whose sequences were determined, 30 contained a common DNA-motif, defining the hexameric consensus sequence CACGTG. We confirm by mutational analysis that binding of the N-Myc derived bHLH-zip domain to this motif is sequence-specific.

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Selected References

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