Sse83871, a useful eight cutter for mammalian genopme analysis (influence of methylation on the activity of Sse83871)

• Published in: Nucleic acids research 1995-07, Vol.23 (13), p.2367-2370
• Main Authors: Sagawa, Hiroaki, Ohshima, Atsushi, Kato, Ikunoshin
• Format: Article
• Language: English
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/23.13.2367

To develop restriction enzymes that are useful for genome analysis, we previously performed screening and isolated Sse8387l from Streptomyces sp. strain 8387. Sse63871 is a restriction enzyme that recognizes 5-CCTGCA/GG-3′ and cleaves DNA at the site shown by the diagonal (Nucleic Acid Res., 18, 5637-5640). The present study evaluated the effects of methylation that is Important when Sse8387l is used for genome analysis. Sse6387l lost cleavage activity after methylation of adenine or methylation of cytosine at any site in the recognition sequence. However, the recognition sequence of Sse6387l contains no CG sequence, which Is the mammalian methylation sequence. In addition, we evaluated the effects of methylation of CG at sites other than the recognition sequence. The cleavage activity of Sse83871 was maintained even when CG sequences were present immediately before or after, or near the recognition sequence, and cytosine was methylated. These results suggest that CG methylation does not affect the cleavage activity of Sse6387l. Therefore, Sse8387l seems to be very useful for mammalian genome analysis.