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Chung,Chung-Nam,Hamaguchi,Yasushi,Honjo,Tasuku,Kawaichi,Masashi 東亞大學校附設遺傳工學硏究所 1995 遺傳工學硏究 Vol.- No.2
To map regions important for DNA binding of the mouse homologue of Suppressor of Hairless or RBP-Jχprotein, mutated mouse RBP-Jχ cDNAs were made by insertion of oligonucleotide linkers or base replacement. DNA binding assays using the mutated proteins expressed in COS cells showed that various mutations between 218 Arg and 227 Arg decreased the DNA binding activity drastically. The DNA binding activity was not affected by amino acid replacements within the integrase motif of the RBP-Jχ protein(230His-269His). Replacements between 291Arg and 323Tyr affected the DNA binding activity slightly but reproducibly. These results indicate that the region encompassing 218Arg-227Arg is critical for the DNA binding activity of RBP-Jχ This region did not show any significant homology to motifs or domains of the previously described DNA binding proteins. Using a truncation mutant protein RBP-Jχ was shown to associate with DNA as a monomer.