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Topic: Methylase

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Restriction enzyme

	Method for cloning and expression of MlyI restriction endonuclease and MlyI methylase and BstNBII methylase in E. coli ...
		Modification methylases recognize and bind to the same recognition sequence as the corresponding restriction endonuclease, but instead of cleaving the DNA, they chemically modify one particular nucleotide within the sequence by the addition of a methyl group (C5 methyl cytosine, N4 methyl cytosine, or N6 methyl adenine).
		Because purified restriction endonucleases and modification methylases are useful tools for creating recombinant molecules in the laboratory, there is a strong commercial interest in obtaining bacterial strains through recombinant DNA techniques that produce large quantities of restriction enzymes.
		A methylase gene with high homology to amino-methyltransferases (N6-adenine methylases) was found in the Micrococcus lylae DNA library after methylase selection.
	www.freepatentsonline.com /6395531.html (3143 words)

	Dam References
		Brooks,J.E., Blumenthal,R.M. and Gingeras,T.R. (1983) The isolation and characterization of the Escherichia coli DNA adenine methylase (dam) gene.
		Herman,G.E. and Modrich,P. (1982) Escherichia coli dam methylase.
		Ritchie,L., Podger,D.M. and Hall,R.M. (1988) A mutation in the DNA adenine methylase gene (dam) of Salmonella typhimurium decreases susceptibility to 9-aminoacridine-induced frameshift mutagenesis.
	users.umassmed.edu /martin.marinus/damrefs.htm (7503 words)

	Method for cloning and expression of PleI restriction endonuclease and PleI and BstNBII methylases in E. coli - Patent ...
		A methylase gene with high homology to amino-methyltransferases (N6-adenine methylases) was found in the Pseudomonas lemoignei DNA library after methylase selection.
		Since methylase selection failed to yield a PleI endonuclease clone, inverse PCR was employed to amplify the adjacent downstream DNA sequence.
		Methylase selection on the Bacillus stearothermophilus 33M genomic DNA had yielded a N6-adenine methylase that conferred protection against PleI digestion when expressed in the pSXY20 plasmid.
	www.freepatentsonline.com /6391608.html (3295 words)

	Proceedings of the Oklahoma Academy of Science
		Although a loss or reduction of some specific methylase (s) in the crude extract from various stages could account for the results thus far discussed, the analysis is complicated by the observation that, after reaching saturation levels, the measured methylating capacity decreases.
		A similar pattern has been shown to represent a methylase inhibitor (6), the activity of which is dependent on the absolute concentration.
		A specific methylase inhibitor in our system was not detected by the assay method of Sharma and Borek (7), but this by no means eliminates the possibility that one (or more) exists.
	digital.library.okstate.edu /oas/oas_htm_files/v57/p16_18.html (1179 words)

	Calzyme - Manufacturers of Enzymes , Proteins , Coenzymes , Substrates and Related Biochemicals (Site not responding. Last check: 2007-11-06)
		Protein methylase II methylates the free carboxyl groups of dicarboxylic acid residues in a protein molecule.
		This pH is favorable for the assay of protein methylase II as interference from other protein methylases is minimal under these experimental conditions (Protein Methylation, Paik, W.K. and Kim, S., eds; Vol I, 113, 1980, Wiley, New York).
		The difference in the radioactivity between the heated and unheated tubes is taken as the radioactivity due to protein-methyl ester formed by the protein methylase II enzyme.
	www.calzyme.com /commerce/catalog/spcategory.jsp?category_id=1091 (493 words)

	Cloning and Characterization of the Lactococcal Plasmid-Encoded Type II Restriction/Modification System, LlaDII -- ...
		The endonuclease gene of 543 bp preceded the methylase gene of
		Nucleotide sequence of the 2,355-bp PstI-EcoRI fragment from plasmid pHW393 and the deduced amino acid sequences of the LlaDII endonuclease and methylase.
		Cloning the modification methylase gene of Bacillus sphaericus R in Escherichia coli.
	aem.asm.org /cgi/content/full/64/7/2424 (4461 words)

	[No title]
		Maekawa, Y., Kawakami, F., Yasukawa, H. Cloning and expression of the BanI restriction endonuclease and DNA methylase genes of Bacillus aneurinolyticus.
		Matvienko, N.N., Kramarov, V.M., Zheleznaya, L.A., Matvienko, N.I. Isolation of site-specific endonuclease and methylase from Bacillus cereus 83.
		Nwosu, V.U., Connolly, B.A., Halford, S.E., Garnett, J. The cloning, purification and characterization of the EcoRV modification methylase.
	www.ibiblio.org /pub/academic/biology/molbio/data/rebase/type2ref.512 (14105 words)

	BCH5425 Molecular Biology and Biotechnology (Site not responding. Last check: 2007-11-06)
		The recognition sequences of the methylases are the same as the associated endonucleases (e.g.
		All methylases transfer the methyl group from S-adenosylmethionine (SAM) to a specific base in the recognition sequence, and SAM is an required component in the methylation reaction.
		The methylase encoded by the dcm gene (dcm methylase) methylates the internal cytosine base, at the C
	wine1.sb.fsu.edu /bch5425/lect04/lect04.htm (1379 words)

	MCB411
		The methylase is a very simple-minded machine--it wanders along DNA and covalently modifies adenines in the sequence -GATC- whenever it finds them.
		One important thing to note at this point is that this is a well-chosen sequence for the task of marking strands--its complement (the sequence on the other strand) is also -GATC- (don't forget that the strands are complementary and anti-parallel).
		This situation won't persist forever--the dam methylase is always chugging along--but it creates a 'window of opportunity' for repair machinery to spot and fix basepairing errors that slipped past the proofreading function of DNA polymerase.
	www.blc.arizona.edu /courses/181lab/MoBiByMe/damRepairNew.html (1172 words)

	cDNA cloning of Batis maritima methyl chloride transferase and purification of the enzyme -- Ni and Hager 95 (22): ...
		methylase defines a new family of plant methyl transferases.
		The lack of catalytic activity of the Batis maritima enzyme in catalyzing the formation of methanethiol is in sharp contrast
		An obvious function for a halophytic methylase would be the maintenance of homeostatic levels of cytoplasmic chloride ion.
	www.pnas.org /cgi/content/full/95/22/12866 (3614 words)

	BCH5425 Molecular Biology and Biotechnology (Site not responding. Last check: 2007-11-06)
		Thus, this combination of a specific methylase and endonuclease functioned as a type of immune system for individual bacterial strains, protecting them from infection by foreign DNA (e.g.
		Of course, this type of protective system is beaten if the attacking phage was previously grown on the same strain as that which it is infecting.
		Structural and biochemical studies have indicated that for the common R/M systems (so called type II), the methylase recognizes and methylates one strand of the DNA duplex, whereas the restriction endonuclease recognizes both strands of the DNA (i.e.
	wine1.sb.fsu.edu /bch5425/lect03/lect03.htm (1200 words)

	Envelope instability in DNA adenine methylase mutants of Salmonella enterica -- Pucciarelli et al. 148 (4): 1171 -- ...
		Envelope instability in DNA adenine methylase mutants of Salmonella enterica -- Pucciarelli et al.
		The percentage of lysis was estimated as 2–5% upon comparison of this signal with that obtained from whole-cell protein extracts (not shown).
		Torreblanca, J. and Casadesus, J. DNA adenine methylase mutants of Salmonella typhimurium and a novel dam-regulated locus.
	mic.sgmjournals.org /cgi/content/full/148/4/1171 (5148 words)

	Salmonella DNA Adenine Methylase Mutants Confer Cross-Protective Immunity -- Heithoff et al. 69 (11): 6725 -- Infection ...
		Badie, G., Heithoff, D. M., Mahan, M. LcrV Synthesis Is Altered by DNA Adenine Methylase Overproduction in Yersinia pseudotuberculosis and Is Required To Confer Immunity in Vaccinated Hosts.
		Chen, L., Paulsen, D. B., Scruggs, D. W., Banes, M. M., Reeks, B. Y., Lawrence, M. Alteration of DNA adenine methylase (Dam) activity in Pasteurella multocida causes increased spontaneous mutation frequency and attenuation in mice.
		Pucciarelli, M. G., Prieto, A. I., Casadesus, J., Garcia-del Portillo, F. Envelope instability in DNA adenine methylase mutants of Salmonella enterica.
	iai.asm.org /cgi/content/abstract/69/11/6725 (744 words)

	Phenotypic and genotypic variation in methylases involved in type II restriction-modification systems in Helicobacter ...
		Phenotypic and genotypic variation in methylases involved in type II restriction-modification systems in Helicobacter pylori -- Takata et al.
		or absence of methylase genes; analysis of the sequenced genomes
		the hypothesis that the acquisition of the methylases by H.pylori
	nar.oxfordjournals.org /cgi/content/full/30/11/2444 (4215 words)

	Restriction Modification System
		Methylase - an enzyme that adds a methyl group to a molecule; in restriction-modification systems of bacteria a methyl group is added to DNA at a specific site to protect the site from restriction endonuclease cleavage
		Several different types of restriction enzymes have been found but the most useful ones for molecular biology and genetic engineering are the Type II restriction enzymes.
		Thus, the viral DNA is restricted in the bacterial cell by the restriction enzyme, and the bacterial DNA is modified by the methylase and is provided protection from its own restriction enzyme.
	www.ndsu.nodak.edu /instruct/mcclean/plsc731/dna/dna5.htm (749 words)

	InterPro: IPR002052 N-6 Adenine-specific DNA methylase
		N-6 adenine-specific DNA methylases (2.1.1.72) (A-Mtase) are enzymes that specifically methylate the amino group at the C-6 position of adenines in DNA.
		Such enzymes are found in the three existing types of bacterial` restriction-modification systems (in type I system the A-Mtase is the product of the hsdM gene, and in type III it is the product of the mod gene).
		P04392 DNA adenine methylase (EC 2.1.1.72) (Deoxyadenosyl-methyltransferase) (M.EcoT4Dam)
	www.ebi.ac.uk /interpro/IEntry?ac=IPR002052 (3200 words)

	Characterization of DNA binding activities of over-expressed KpnI restriction endonuclease and modification methylase
		Even though the methylase clone is in a low copy number plasmid pACMK, high level expression of methylase is achieved.
		The yield of purified endonuclease and methylase from one liter of culture was approximately 30 and 6 mg respectively.
		In contrast to KpnI methylase cloning, Enase gene was cloned as a promoter-less fragment into two separate expression vectors.
	www.ias.ac.in /jbiosci/september1999/article3.htm (4597 words)

	HaeIII Methylase, Methylases, New England Biolabs
		One unit is defined as the amount of enzyme required to protect 1 µg λ DNA in 1 hour at 37°C in a total reaction volume of 10 µl against cleavage by HaeIII restriction endonuclease.
		A 50 μl reaction containing 1 μg of λDNA (HindIII digest) and 100 units of HaeIII Methylase incubated for 16 hours at 37ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
		Incubation of a 50 μl reaction containing 100 units of HaeIII Methylase with 1 μg of ΦX174 RF I DNA for 4 hours at 37ºC resulted in
	www.neb.com /nebecomm/products/productM0224.asp (317 words)

	Hot Topics! Rb-mediated Transcriptional Repression by a Histone Methylase Complex
		The retinoblastoma gene (Rb) is a master regulator of the cell cycle, capable of silencing target genes that mediate cell entry into the S phase.
		It has recently been demonstrated that the mammalian histone methylase SUV39H1 (specific for lysine 9 of histone H3) and the associated methyl-lysine binding protein HP1 can repress gene expression.
		Nielsen and colleagues now demonstrate that the Rb protein binds to SUV39H1/HP1 and in doing so, recruits the complex to the E2F/Rb-regulated gene promoters to mediate repression.
	www.ambion.com /techlib/hottopics/rb_082701.html (441 words)

	AluI Methylase, Methylases, New England Biolabs
		One unit is defined as the amount of enzyme required to protect 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 10 µl against cleavage by AluI restriction endonuclease.
		A 50 μl reaction containing 1 μg of λDNA and 40 units of AluI Methylase incubated for 16 hours at 37ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
		Incubation of a 50 μl reaction containing 40 units of AluI Methylase with 1 μg of a mixture of single and double-stranded [
	www.neb.com /nebecomm/products/productM0220.asp (291 words)

	Characterization of an Extremely Thermostable Restriction Enzyme, PspGI, from a Pyrococcus Strain and Cloning of the ...
		and 10 were found among the cytosine-5 methylases (20).
		The recognition sequence of PspGI is the same as that of the Dcm methylase.
		The Dcm methylase modifies the internal C at the
	aem.asm.org /cgi/content/full/64/10/3669 (3635 words)

	Functional analysis of putative restriction-modification system genes in the Helicobacter pylori J99 genome -- Kong et ...
		Dot blot assay to measure methylase activities using rabbit primary antibodies that react specifically with DNA containing m6A or m4C modifications.
		(A) Methylase dot blot assay using m6A antibody with dilution of 1:750 000.
		both endonuclease and methylase genes have to be turned off.
	nar.oxfordjournals.org /cgi/content/full/28/17/3216 (4853 words)

	DNA Adenine Methylase Is Essential for Viability and Plays a Role in the Pathogenesis of Yersinia pseudotuberculosis ...
		DNA Adenine Methylase Is Essential for Viability and Plays a Role in the Pathogenesis of Yersinia pseudotuberculosis and Vibrio cholerae -- Julio et al.
		DNA Adenine Methylase Is Essential for Viability and Plays a Role in the Pathogenesis of Yersinia pseudotuberculosis and Vibrio cholerae
		Yang, J. M., DeUrraza, P. J., Matvienko, N., O'Sullivan, D. Involvement of the LlaKR2I Methylase in Expression of the AbiR Bacteriophage Defense System in Lactococcus lactis subsp.
	iai.asm.org /cgi/content/abstract/69/12/7610 (754 words)

	Plasmid-Mediated 16S rRNA Methylase in Serratia marcescens Conferring High-Level Resistance to Aminoglycosides -- Doi ...
		Plasmid-Mediated 16S rRNA Methylase in Serratia marcescens Conferring High-Level Resistance to Aminoglycosides -- Doi et al.
		Park, Y.-J., Lee, S., Yu, J. K., Woo, G.-J., Lee, K., Arakawa, Y. Co-production of 16S rRNA methylases and extended-spectrum {beta}-lactamases in AmpC-producing Enterobacter cloacae, Citrobacter freundii and Serratia marcescens in Korea.
		Yan, J.-J., Wu, J.-J., Ko, W.-C., Tsai, S.-H., Chuang, C.-L., Wu, H.-M., Lu, Y.-J., Li, J.-D. Plasmid-mediated 16S rRNA methylases conferring high-level aminoglycoside resistance in Escherichia coli and Klebsiella pneumoniae isolates from two Taiwanese hospitals.
	aac.asm.org /cgi/content/abstract/48/2/491 (574 words)

	Erythromycin-Resistant Neisseria gonorrhoeae and Oral Commensal Neisseria spp. Carry Known rRNA Methylase Genes -- ...
		Carry Known rRNA Methylase Genes -- Roberts et al.
		The methylase gene composition of oral commensal Neisseria spp.
		Mobility of rRNA methylase genes in Campylobacter (Wolinella) rectus.
	aac.asm.org /cgi/content/full/43/6/1367 (3370 words)

	SIUC COS Microbiology 421 - chapter 12
		Methylase gene should be expressed before the restriction enzyme
		The restriction enzyme should have a leader peptide so that it will be transported from the cytoplasm to the periplasm
		Target genes encode both th methylase and the restriction enzyme
	www.science.siu.edu /microbiology/micr421/chapter12.html (600 words)

	EC 2.1.1.37
		Kalousek, F. and Morris, N.R. The purification and properties of deoxyribonucleic acid methylase from rat spleen.
		Roy, P.H. and Weissbach, A. DNA methylase from HeLa cell nuclei.
		Turnbull, J.F. and Adams, R.L.P. DNA methylase: purification from ascites cells and the effect of various DNA substrates on its activity.
	www.chem.qmul.ac.uk /iubmb/enzyme/EC2/1/1/37.html (166 words)

	BB331 Lecture 8
		Newly synthesized strands during DNA replication are temporarily (that is, for a short period of time) unmethylated, and DNA with one strand methylated and the other strand unmethylated is called hemi-methylated.
		Fully unmethylated DNA only occurs in cells lacking dam methylase activity, and those cells have increased rates of mutation (mutator phenotype) and increased recombination frequencies.
		Endonuclease that cleaves unmethylated strand just 5' to the G in the sequence GATC (that is, N
	oregonstate.edu /instruction/bb331/lecture08/lecture08.html (1229 words)

	Quantitative Analysis of EcoR1 Methylase-DNA Complex by Atomic Force Microscopy (Site not responding. Last check: 2007-11-06)
		We directly applied atomic force microscopy (AFM) to investigate linear pBR322-EcoR1 methylase complexes and quantitatively analyzed the bend angles of linear pBR322-EcoR1 methylase complexes and the bound protein widths.
		The data indicated that the unacute angle state was the result of unique interactions between EcoR1 methylase and the recognition site and the complementary site on duplex DNAs, and suggested that the acute angle conformation could be an intermediate in the formation of the unacute angle state.
		Our works provide a detail insight into the DNA structural variations involved in EcoR1 methylase-binding processes and demonstrate further the versatility of AFM as an imaging technique for studying the interaction between large DNA fragment and protein.
	www.scanning-fams.org /scanabstracts/SCANNING06/28015.html (210 words)

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