{"id":1764,"date":"2022-04-04T22:03:00","date_gmt":"2022-04-04T15:03:00","guid":{"rendered":"http:\/\/sibenzyme.com\/product\/kro-i\/"},"modified":"2026-01-11T17:58:00","modified_gmt":"2026-01-11T10:58:00","slug":"kro-i","status":"publish","type":"product","link":"https:\/\/sibenzyme.com\/en\/product\/kro-i\/","title":{"rendered":"Kro I"},"content":{"rendered":"

Recognition site and hydrolysis position<\/strong>:<\/p>\n\n\n\n\n
GC(5mC)GGC<\/td>\nG\u2191C(5mC)GGC<\/td>\n<\/tr>\n
CGG(5mC)CG<\/td>\nCGG(5mC)C\u2193G<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Source:<\/strong> Kocurea rosea 307
Substrate specificity<\/strong>:
Unit definition: <\/strong>One unit is defined as the amount of enzyme required to hydrolyze completely 1 \u03bcg of linearized plasmid pMHpaII 1 in 1 hour at 37\u00b0C in a total reaction volume of 50 \u03bcl.<\/p>\n\n\n\n
KroI activity assay on DNA pMHpaII 1\/DriI <\/b>
Lanes:
1 and 6 \u2013 1 Kb SE DNA-markers.
2 \u2013 Control DNA pMHpaII 1\/DriI,
3 \u2013 0.5 \u03bcl Kro I
4 \u2013 1 \u03bcl Kro I
5 \u2013 2 \u03bcl Kro I
Products were separated in 1,4% agarose gel in Buffer TAE.<\/td>\n
\"KroI<\/center><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Assayed on: <\/strong>DNA pMHpaII 1\/DriI is a linearized plasmid pMHpaII 1. pMHpaII 1 carries a gene of DNA-methyltransferase M.HpaII, which methylates sites 5`-CCGG-3` producing 5`-C(5mC)GG-3`\/3`-GG(5mC)C-5`, and includes three canonical sites
5`-GC(5mC)GGC-3`\/3`-CGG(5mC)CG-5`.
Optimal SE-buffer:<\/strong> SE-buffer G
Enzyme activity (%)<\/strong>:<\/p>\n\n\n\n\n
B<\/td>\nG<\/td>\nO<\/td>\nW<\/td>\nY<\/td>\nROSE<\/td>\n<\/tr>\n
50<\/td>\n100<\/td>\n25<\/td>\n50<\/td>\n75<\/td>\n100<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Storage conditions<\/strong>: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 \u03bcg \/ml BSA; 50% glycerol; Store at -20\u00b0C.
Ligation<\/strong>: –
Non-specific hydrolisis<\/strong>: No detectable degradation of 1\u03bcg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 30\u00b0C in a total reaction volume of 50 \u03bcl.Reagents Supplied with Enzyme<\/b>
10 X SE-buffer G
Methylation sensitivity<\/b>: The enzyme cleaves C5-methylated DNA and doesn’t cut unmodified DNA [1]<\/a>.
Notes: <\/strong>References<\/b>:<\/strong>
1. Dedkov V.S., Rechkunova N.I., Degtyarev S.Kh., Zhilkin P.A., Kolykhalov A.A., Verkhozina V.A. Bacillus sphaericus – a strain producer of restriction endonuclease BsiI.\/\/ Soviet Union patent SU 1784642 (1992). (In Russian)
2. Chernukhin V.A., Zhuravleva R.O., Tarasova G.V., Boltengagen A. A., Akishev A.G., Mikhnenkova N.A., Degtyarev S.Kh.<\/i><\/b> Bacterial strain Kocuria rosea – producer of KroI site specific endonuclease.<\/b><\/span> \/\/ Russian Federation patent RU 2394099 C1 (2010).
 
1. V.A. Chernukhin, E.V. Kileva, J.E. Tomilova, A.A. Boltengagen, V.S. Dedkov, N.A. Mikhnenkova, D.A. Gonchar, L.N. Golikova, S.Kh. Degtyarev A new methyl-directed site-specific endonuclease KroI recognizes and cuts DNA sequence 5\u2019-G^C(5mC)GGC-3\u2019<\/a> \/\/ Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.7, No 1, pp 14-20, 2011
2. Chernukhin V.A., Zhuravleva R.O., Tarasova G.V., Boltengagen A. A., Akishev A.G., Mikhnenkova N.A., Degtyarev S.Kh.<\/b><\/i> Bacterial strain Kocuria rosea – producer of KroI site specific endonuclease.<\/b><\/font> \/\/ Russian Federation patent RU 2394099 C1 (2010).<\/p>","protected":false},"excerpt":{"rendered":"

MD DNA endonuclease Kro 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-1764","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-4","9":"pa_4ebaafd1-51f465affc","10":"pa_50b54da1-pa_50b54da_6e490e","11":"pa_be110261-pa_be11026_383772-2","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-afbfdccfa7","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\/1764","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=1764"}],"wp:term":[{"taxonomy":"product_brand","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_brand?post=1764"},{"taxonomy":"product_cat","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_cat?post=1764"},{"taxonomy":"product_tag","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_tag?post=1764"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}