{"id":1770,"date":"2022-04-04T22:03:00","date_gmt":"2022-04-04T15:03:00","guid":{"rendered":"http:\/\/sibenzyme.com\/product\/pkr-i\/"},"modified":"2026-01-11T17:58:01","modified_gmt":"2026-01-11T10:58:01","slug":"pkr-i","status":"publish","type":"product","link":"https:\/\/sibenzyme.com\/en\/product\/pkr-i\/","title":{"rendered":"Pkr I"},"content":{"rendered":"

Recognition site and hydrolysis position<\/strong>:<\/p>\n\n\n\n\n\n
DNA sequence with at least three 5mC: <\/td>\nDNA sequence with at least three 5mC: <\/td>\n<\/tr>\n
GCN\u2191GC<\/td>\nGCN\u2191GC<\/td>\n<\/tr>\n
CG\u2193NCG<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Source:<\/strong> Planomicrobium koreense 78k
Substrate specificity<\/strong>:
Unit definition: <\/strong>One unit is defined as the amount of enzyme
required to hydrolyze completely 1 μg of linearized plasmid pFsp4HI3 in 1 hour at 37°C in a total reaction volume of 50 μl (see run 4 in the figure). <\/p>\n\n\n\n
PkrI 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 Pkr I
4 \u2013 1 μl Pkr I
5 \u2013 2 μl Pkr I <\/p>\n

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

PkrI 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 three sites:
5`-G(5mC)NG(5mC)-3`\/3`-CGN(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
50<\/td>\n75<\/td>\n10<\/td>\n25<\/td>\n100<\/td>\n100<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Storage conditions<\/strong>: 20 mM Tris-HCl (pH 7.4); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 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 2 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 DNA and doesn’t cut unmodified DNA [1]<\/a>.
Notes: <\/strong>References<\/b>:<\/strong>
1.Degtyarev S. Kh., Abdurashitov M.A., Kolyhalov A.A., Rechkunova N.I. Acl I, a new restriction endonuclease from Acinetobacter calcoaceticus recognizing 5'-AA^CGTT -3'. \/\/ Nucleic Acids Research, Vol. 20, No. 14, 37-87 (1992)
2. Chmuzh E.V., Kashirina Yu.G., Tomilova Yu.E., Chernukhin V.A., Okhapkina S.S., Gonchar D. A., Dedkov V.S., Abdurashitov M. A., Degtyarev S. Kh.<\/b><\/i> Gene cloning, comparative analysis of the protein structures from Fsp4HI restriction-modification system and biochemical characterization of the recombinant DNA methyltransferase M.Fsp4HI.<\/b><\/font> \/\/ Molecular Biology, V.41, No 1, p. 43-50 (2007)<\/p>\n

1.V.A. Chernukhin, T. N. Nayakshina, D.A. Gonchar, J.E. Tomilova, M.V. Tarasova, V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtyarev A new site-specific methyl-directed DNA endonuclease PkrI recognizes and cuts methylated DNA sequence 5′-GCN^GC-3’\/3′-CG^NCG-5′ carrying at least three 5-methylcytosines<\/a> \/\/ Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.7, No 3, pp 35-42, 2011
2. Chmuzh E.V., Kashirina Yu.G., Tomilova Yu.E., Chernukhin V.A., Okhapkina S.S., Gonchar D. A., Dedkov V.S., Abdurashitov M. A., Degtyarev S. Kh.<\/b><\/i> Gene cloning, comparative analysis of the protein structures from Fsp4HI restriction-modification system and biochemical characterization of the recombinant DNA methyltransferase M.Fsp4HI.<\/b><\/font> \/\/ Molecular Biology, V.41, No 1, p. 43-50 (2007)
<\/p>","protected":false},"excerpt":{"rendered":"

MD DNA endonuclease Pkr 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-1770","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_4ebaafd1-8ccf0454b7","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-b4910ec8ea","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\/1770","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=1770"}],"wp:term":[{"taxonomy":"product_brand","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_brand?post=1770"},{"taxonomy":"product_cat","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_cat?post=1770"},{"taxonomy":"product_tag","embeddable":true,"href":"https:\/\/sibenzyme.com\/en\/wp-json\/wp\/v2\/product_tag?post=1770"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}