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Three proteins, yeast transcription regulatory protein GCN4, M FokI DNA methyltransferase and R FokI restriction endonuclease (ENase) were used to attain specific cleavage of DNA at the 18-20-bp GCN4 recognition site. This is a novel version of the Achilles heel cleavage (AC) technique [Koob et al., Science 241 (1988) 1084-1086]. Since the method employs a class-IIS ENase (R FokI), which...
SacNI, an isoschizomer of the restriction endonuclease, BanII [Sugisaki et al., Nucleic Acids Res. 10 (1982) 5747-5752], has been isolated from Streptomyces achromogenes N-J-H. SacNI recognizes the palindromic sequence, 5 -GRGCY/C, and cleaves within the recognition sequence, generating a 3 protruding RGCY end (where R = A or G, and Y = C or G).
The restriction endonuclease AvcI, an isoschizomer of Sau96I [Sussenbach et al., Nucleic Acids Res. 5 (1978) 1153-1163], was purified from Actinomyces cristalomycini. AvcI recognizes a 5-bp palindromic sequence, 5 -G↓GNCC and cleaves it after the first G residue producing a 3-nucleotide 5 -overhang.
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