Determination of Methylation Site of DNA-Methyltransferase NlaX by a Hybrid Method. (September 2002)

• Record Type: Journal Article
• Authors: Kubareva, E.A.; Walter, J.; Karyagina, A.S.; Vorob'eva, O.V.; Lau, P.C.K.; Trautner, T.
• Is Part Of: Biotechniques. Volume 33:Number 3(2002)
• Page Start: 526
• Page End: 531
• Publication Date: 2002-09
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/02333st02_11839a
• Languages: English
• ISSNs: 0736-6205

Abstract

Using a new method based on a combination of bisulfite reaction, the repair enzyme uracil-DNA glycosylase, and synthetic oligodeoxyribonucleotides, the methylation site of DNA-methyltransferase Nla X (M· Nla X) from Neisseria lactamica was established to be the inner cytosine in the double-stranded pentanucleotide recognition sequence 5′-CCNGG-3′ (where N = any nucleoside). 5-Methylcytosine (m 5 C) type modification by M· Nla X was confirmed by the use of oligonucleotide substrates that contain 5-fluoro-2′-deoxycytidine.