Summary
Restriction endonuclease R from Bacillus subtilis strain R cleaves nonmodified SPP1 DNA in approximately 80, and λ DNA in about 200 different sites. DNA digests with this endonuclease and with endonuclease Hae III from Haemophilus aegyptius show identical fragmentation patterns on gel electrophoresis, indicating that the two enzymes recognise the same nucleotide sequence. The polynucleotide kinase reaction was used in conjunction with two-dimensional ionophoretic nucleotide mapping methods to identify the 5′-nucleotide sequences at the sites of cleavage by the B. subtilis restriction endonuclease.
The results show that the recognition sequence is \(\begin{gathered} 5'{\text{ }}{\text{N}}{\text{G}}{\text{G}} \downarrow \user2{C}\user2{C}{\text{N}}\user2{3'} \hfill \\ \user2{3'}{\text{ }}{\text{N}}{\text{G}}{\text{G}} \uparrow \user2{C}\user2{C}{\text{N}}5' \hfill \\ \end{gathered} \) where arrows indicate the points of strand scission. Each of the four possible nucleotides can occur in the positions flanking the recognition site.
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Bron, S., Murray, K. Restriction and modification in B. subtilis . Molec. Gen. Genet. 143, 25–33 (1975). https://doi.org/10.1007/BF00269417
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DOI: https://doi.org/10.1007/BF00269417