{"id":1752,"date":"2022-04-04T22:03:00","date_gmt":"2022-04-04T15:03:00","guid":{"rendered":"http:\/\/sibenzyme.com\/product\/glu-i\/"},"modified":"2026-01-11T17:58:00","modified_gmt":"2026-01-11T10:58:00","slug":"glu-i","status":"publish","type":"product","link":"https:\/\/sibenzyme.com\/en\/product\/glu-i\/","title":{"rendered":"Glu I"},"content":{"rendered":"

Recognition site and hydrolysis position<\/strong>:<\/p>\n\n\n\n\n
G(5mC)NG(5mC)<\/td>\nG(5mC)\u2191NG(5mC)<\/td>\n<\/tr>\n
(5mC)GN(5mC)G<\/td>\n(5mC)GN\u2193(5mC)G<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Source:<\/strong> Glacial ice bacterium GL24
Substrate specificity<\/strong>:
Unit definition: <\/strong>One unit is defined as the amount of enzyme
required to hydrolyze completely a unique site:
5`-G(5mC)NG(5mC)-3`\/3`-(5mC)GN(5mC)G-5`
in 1 μg of linearized plasmid pFsp4HI3\/DriI in 1 hour at 37°C in a total reaction volume of 50 μl. As a result a linearized plasmid pFsp4HI3 disappears (see run 4 in the figure). GluI digestion of recognition sequences with three 5-methylcytosines results in additional bands appearance (runs 4-6 in the figure). <\/p>\n\n\n\n
GluI activity assay on DNA pFsp4HI3\/DriI <\/b>
Lanes:
1 and 6 \u2013 1 Kb SE DNA-markers.
2 \u2013 Control DNA pFsp4HI3\/DriI,
3 \u2013 0.5 μl Glu I
4 \u2013 1 μl Glu I
5 \u2013 2 μl Glu I <\/p>\n

Products were separated in 1,4% agarose gel in Buffer TAE.<\/td>\n

GluI activity assay on DNA pFsp4HI3\/DriI<\/center><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Assayed on: <\/strong>DNA pFsp4HI3\/DriI is a linearized plasmid pFsp4HI3, which carries a gene of DNA-methyltransferase M.Fsp4HI and includes a unique GluI site:5`-G(5mC)NG(5mC)-3`\/3`-(5mC)GN(5mC)G-5` [2]<\/a>.
Optimal SE-buffer:<\/strong> SE-buffer Y
Enzyme activity (%)<\/strong>:<\/p>\n\n\n\n\n\n
B<\/td>\nG<\/td>\nO<\/td>\nW<\/td>\nY<\/td>\nROSE<\/td>\n<\/tr>\n
75<\/td>\n75<\/td>\n25<\/td>\n50<\/td>\n100<\/td>\n100<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Storage conditions<\/strong>: 10 mM KH2<\/sub>PO4<\/sub> (pH 7.45); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg\/ml BSA, 50% glycerol; Store at -20°C.
Ligation<\/strong>: –
Non-specific hydrolisis<\/strong>: No detectable degradation of 1μg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 37°C in a total reaction volume of 50 μl.<\/td>\n<\/tr>\n

Reagents Supplied with Enzyme<\/b><\/td>\n10 X SE-buffer Y
Methylation sensitivity<\/b>: The enzyme cleaves C5-methylated DNAand does not cut unmodified DNA [1]<\/a>.
Notes: <\/strong>When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
References<\/b>:<\/strong>
1. Chernukhin V.A., Chmuzh E.V., Tomilova J.E., Nayakshina T.N., Dedkov V.S., Degtyarev S.Kh.<\/b><\/i> Bacterial strain Glacial ice bacterium – producer of GluI site specific endonuclease. <\/b><\/font>\/\/ Russian Federation patent RU 2322492 C1 (2006).
2. Zhilkina O.A., Repin V.E., Degtyarev S.Kh. Preparation method of restriction endonuclease SfaNI, that recognizes and cleaves DNA sequence 5′-GCATC(N)5^, 3′-CGTAG(N)9^.\/\/ Soviet Union patent SU 1442546 (1988). (In Russian)
Application:<\/b>
Degtyarev S.Kh., Repin V.E., Rechkunova N.I. Proteus vulgaris – a strain producer of restriction endonuclease PvuII.\/\/ Soviet Union patent SU 1632974 (1991). (In Russian)
Kravets AP, Mousseau TA, Litvinchuk AV, Ostermiller Sh, Vengen GS, Grodzinski DM. <\/i><\/b> Changes in wheat DNA methylation pattern after chronic seed gamma-irradiation.<\/b><\/font>\/\/ Tsitol Genet. 2010 Sep-Oct;44(5):18-22. Russian.
1. Chernukhin V.A., Chmuzh E.V., Tomilova J.E., Nayakshina T.N., Dedkov V.S., Degtyarev S.Kh.<\/b><\/i> Bacterial strain Glacial ice bacterium – producer of GluI site specific endonuclease. <\/b><\/font>\/\/ Russian Federation patent RU 2322492 C1 (2006).
2. V.A. Chernukhin, E.V. Chmuzh, Yu.E. Tomilova, T.N. Nayakshina, D.A. Gonchar, V.S. Dedkov, S.Kh. Degtyarev A novel site-specific endonuclease GluI recognizes methylated DNA sequence 5\u2019-G(5mC)^NG(5mC)-3\u2019\/3\u2019-(5mC)GN^(5mC)G.<\/a> \/\/ Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 2, pp 13-17, 2007
Application:<\/b>
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro<\/i> and in silico<\/i><\/a> \/\/ Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 4, pp 19-27, 2007
Kravets AP, Mousseau TA, Litvinchuk AV, Ostermiller Sh, Vengen GS, Grodzinski DM. <\/i><\/b> Changes in wheat DNA methylation pattern after chronic seed gamma-irradiation.<\/b><\/font>\/\/ Tsitol Genet. 2010 Sep-Oct;44(5):18-22. Russian. <\/p>","protected":false},"excerpt":{"rendered":"

MD DNA endonuclease Glu I.
The enzyme cleaves C5-methylated DNA and doesn’t cut unmodified DNA.<\/p>\n","protected":false},"featured_media":58,"template":"","meta":[],"product_brand":[],"product_cat":[69],"product_tag":[921],"class_list":{"0":"post-1752","1":"product","2":"type-product","3":"status-publish","4":"has-post-thumbnail","6":"product_cat-md","7":"product_tag-epigenetika","8":"pa_285b89be-pa_285b89b_2d3edf","9":"pa_4ebaafd1-a930084bac","10":"pa_50b54da1-pa_50b54da_6e490e","11":"pa_be110261-pa_be11026_383772-3","12":"pa_concentration-pa_concent_8cc665-6","13":"pa_fasovka-pa_fasovka_e7d4b7-6","14":"pa_fasovka-pa_fasovka_e7d4b7-7","15":"pa_package-d20caec3b4","16":"pa_package-5dbc98dcc9","17":"pa_site-4e87138aa5","18":"pa_variant-d2203cb123","20":"first","21":"instock","22":"taxable","23":"shipping-taxable","24":"purchasable","25":"product-type-variable","26":"berocket_lgv_grid","27":"berocket_lgv_list_grid"},"_links":{"self":[{"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product\/1752","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product"}],"about":[{"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/types\/product"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/media\/58"}],"wp:attachment":[{"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/media?parent=1752"}],"wp:term":[{"taxonomy":"product_brand","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_brand?post=1752"},{"taxonomy":"product_cat","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_cat?post=1752"},{"taxonomy":"product_tag","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_tag?post=1752"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}