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<h1 id="firstHeading" class="firstHeading mw-first-heading"><span class="mw-page-title-main">FokI</span></h1>
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<div id="mw-content-text" class="mw-body-content mw-content-ltr" lang="de" dir="ltr"><div class="mw-content-ltr mw-parser-output" lang="de" dir="ltr"><table class="wikitable hintergrundfarbe-basis infobox float-right" id="Vorlage_Infobox_Protein_Type-2_restriction_enzyme_FokI" style="font-size:90%; margin-top:0; width:350px;" summary="Infobox Protein">
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<th colspan="3" style="background:#90EE90; color:#202122;">Type-2 restriction enzyme FokI
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<td colspan="3"><span typeof="mw:File"></span>
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<td colspan="3" class="hintergrundfarbe1" style="text-align:center; font-size:smaller; font-weight:bold;">FokI an DNA gebunden, nach <a href="Protein_Data_Bank" title="Protein Data Bank">PDB</a> <a rel="nofollow" class="external text" href="https://www.rcsb.org/structure/1FOK">1FOK</a>
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<td>Andere Namen
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<p>Endonuclease FokI, Type II restriction enzyme FokI, Type IIS restriction enzyme FokI
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<p>Vorhandene Strukturdaten: <a href="Protein_Data_Bank" title="Protein Data Bank">PDB</a> <a rel="nofollow" class="external text" href="https://www.rcsb.org/structure/2FOK">2FOK</a>
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<td><a href="Molare_Masse" title="Molare Masse">Masse</a>/Länge <a href="Prim%C3%A4rstruktur" title="Primärstruktur">Primärstruktur</a>
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<td colspan="2" style="text-align:center;">583 <a href="Aminos%C3%A4uren" title="Aminosäuren">Aminosäuren</a>, 66.219 <a href="Atomare_Masseneinheit" title="Atomare Masseneinheit">Da</a>
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<td><a href="Sekund%C3%A4rstruktur" title="Sekundärstruktur">Sekundär-</a> bis <a href="Quart%C3%A4rstruktur" title="Quartärstruktur">Quartärstruktur</a>
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<td colspan="2" style="text-align:center;"><a href="Homodimer" class="mw-redirect" title="Homodimer">Homodimer</a>
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<th colspan="3" style="background:#90EE90; color:#202122;">Bezeichner
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<td>Externe IDs
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<ul><li><a href="UniProt" title="UniProt">UniProt</a> <a rel="nofollow" class="external text" href="https://www.uniprot.org/uniprotkb/P14870">P14870</a></li></ul>
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<th colspan="3" style="background:#90EE90; color:#202122;">Enzymklassifikation
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<td><a href="EC-Nummer" title="EC-Nummer">EC, Kategorie</a>
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<td colspan="2" class="" style="text-align:center;"><a rel="nofollow" class="external text" href="https://www.brenda-enzymes.org/enzyme.php?ecno=3.1.21.4">3.1.21.4</a>
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</tbody></table><p><span class="editoronly" style="display:none;"></span>
</p><p><b>FokI</b> ist ein <a href="Restriktionsenzym" title="Restriktionsenzym">Restriktionsenzym</a> aus dem Bakterium <i>Flavobacterium okeanokoites</i>.
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<div class="mw-heading mw-heading2"><h2 id="Eigenschaften">Eigenschaften</h2></div>
<p>FokI ist eine <a href="Endonuklease" title="Endonuklease">Endonuklease</a> (Typ II, Subtyp S), die <a href="DsDNA" class="mw-redirect" title="DsDNA">dsDNA</a> nach der Erkennungssequenz 5'-GGATG-3' schneidet, je nach Strang 9 bzw. 13 <a href="Nukleotid" class="mw-redirect" title="Nukleotid">Nukleotide</a> hinter der Erkennungssequenz.<sup id="cite_ref-Wah_1-0" class="reference"><a href="#cite_note-Wah-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> Durch den Schnitt der doppelsträngigen DNA durch FokI entsteht ein <i><a href="Sticky_ends" class="mw-redirect" title="Sticky ends">sticky end</a></i> (Ende mit 4 Nukleotiden Überhang) und je einer <a href="Phosphatgruppe" class="mw-redirect" title="Phosphatgruppe">Phosphatgruppe</a> an beiden 5'-Enden der doppelsträngigen DNA-Produkte. FokI wird durch eine DCM-Methylierung und durch eine <a href="CpG-Dinukleotid" title="CpG-Dinukleotid">CpG</a>-Methylierung gehemmt, nicht aber durch eine <a href="Dam-Methylase" title="Dam-Methylase">DAM-Methylierung</a>. Nach einer Restriktion von DNA <i>in vitro</i> kann FokI durch 20-minütiges Erhitzen auf 65 °C <a href="Denaturierung_(Biochemie)" title="Denaturierung (Biochemie)">denaturiert</a> und somit inaktiviert werden. Meistens wird FokI als <a href="Rekombinantes_Protein" title="Rekombinantes Protein">rekombinantes Protein</a> in <i>E.coli</i> hergestellt. FokI schneidet dsDNA mit zwei Erkennungssequenzen in beliebiger Orientierung besser als mit einer, wobei sie mehrere hundert Nukleotide auseinander liegen können.<sup id="cite_ref-Halford_2-0" class="reference"><a href="#cite_note-Halford-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup>
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<div class="mw-heading mw-heading2"><h2 id="Anwendungen">Anwendungen</h2></div>
<p>FokI wird für <a href="Restriktionsverdau" title="Restriktionsverdau">Restriktionsverdaue</a> im Rahmen von <a href="Klonierung" title="Klonierung">Klonierungen</a> oder <a href="Restriktionskarte" title="Restriktionskarte">Restriktionsanalysen</a> (vor allem von <a href="Polymorphismus" title="Polymorphismus">Polymorphismen</a> des <a href="Gen" title="Gen">Gens</a> des <a href="Vitamin-D-Rezeptor" title="Vitamin-D-Rezeptor">Vitamin-D-Rezeptors</a> zur Bestimmung der Anfälligkeit für <a href="Tuberkulose" title="Tuberkulose">Tuberkulose</a><sup id="cite_ref-3" class="reference"><a href="#cite_note-3"><span class="cite-bracket">[</span>3<span class="cite-bracket">]</span></a></sup> und <a href="Nierenversagen" title="Nierenversagen">Nierenversagen</a><sup id="cite_ref-4" class="reference"><a href="#cite_note-4"><span class="cite-bracket">[</span>4<span class="cite-bracket">]</span></a></sup>) verwendet. Der enzymatische Anteil von FokI wird im Zuge eines <a href="Proteindesign" title="Proteindesign">Proteindesigns</a> mit den Erkennungsdomänen anderer Restriktionsenzyme als <a href="Fusionsprotein" title="Fusionsprotein">Fusionsprotein</a> verwendet, z. B. zur Erzeugung von <a href="Zinkfingernuklease" class="mw-redirect" title="Zinkfingernuklease">Zinkfingernukleasen</a><sup id="cite_ref-Halford_2-1" class="reference"><a href="#cite_note-Halford-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup> oder in Kombination mit einem inaktivierten <a href="Cas9" title="Cas9">Cas9</a>.<sup id="cite_ref-5" class="reference"><a href="#cite_note-5"><span class="cite-bracket">[</span>5<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-6" class="reference"><a href="#cite_note-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-7" class="reference"><a href="#cite_note-7"><span class="cite-bracket">[</span>7<span class="cite-bracket">]</span></a></sup>
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<div class="mw-heading mw-heading2"><h2 id="Einzelnachweise">Einzelnachweise</h2></div>
<ol class="references">
<li id="cite_note-Wah-1"><span class="mw-cite-backlink"><a href="#cite_ref-Wah_1-0">↑</a></span> <span class="reference-text">D. A. Wah, J. Bitinaite, I. Schildkraut, A. K. Aggarwal: <i>Structure of FokI has implications for DNA cleavage.</i> In: <i><a href="Proceedings_of_the_National_Academy_of_Sciences" class="mw-redirect" title="Proceedings of the National Academy of Sciences">Proceedings of the National Academy of Sciences</a>.</i> Band 95, Nummer 18, September 1998, S. 10564–10569, <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/9724743?dopt=Abstract">PMID 9724743</a>, <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC27934/">PMC 27934</a> (freier Volltext).</span>
</li>
<li id="cite_note-Halford-2"><span class="mw-cite-backlink">↑ <sup><a href="#cite_ref-Halford_2-0">a</a></sup> <sup><a href="#cite_ref-Halford_2-1">b</a></sup></span> <span class="reference-text">S. E. Halford, L. E. Catto, C. Pernstich, D. A. Rusling, K. L. Sanders: <i>The reaction mechanism of FokI excludes the possibility of targeting zinc finger nucleases to unique DNA sites.</i> In: <i>Biochemical Society transactions.</i> Band 39, Nummer 2, April 2011, S. 584–588, <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1042/BST0390584">10.1042/BST0390584</a></span>, <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/21428944?dopt=Abstract">PMID 21428944</a>.</span>
</li>
<li id="cite_note-3"><span class="mw-cite-backlink"><a href="#cite_ref-3">↑</a></span> <span class="reference-text">Y. Cao, X. Wang, Z. Cao, X. Cheng: <i>Vitamin D receptor gene FokI polymorphisms and tuberculosis susceptibility: a meta-analysis.</i> In: <i>Archives of Medical Science.</i> Band 12, Nummer 5, Oktober 2016, S. 1118–1134, <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.5114/aoms.2016.60092">10.5114/aoms.2016.60092</a></span>, <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/27695504?dopt=Abstract">PMID 27695504</a>, <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5016579/">PMC 5016579</a> (freier Volltext).</span>
</li>
<li id="cite_note-4"><span class="mw-cite-backlink"><a href="#cite_ref-4">↑</a></span> <span class="reference-text">L. Yang, L. Wu, Y. Fan, J. Ma: <i>Associations among four polymorphisms (BsmI, FokI, TaqI and ApaI) of vitamin D receptor gene and end-stage renal disease: a meta-analysis.</i> In: <i>Archives of medical research.</i> Band 46, Nummer 1, Januar 2015, S. 1–7, <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1016/j.arcmed.2014.11.017">10.1016/j.arcmed.2014.11.017</a></span>, <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/25434518?dopt=Abstract">PMID 25434518</a>.</span>
</li>
<li id="cite_note-5"><span class="mw-cite-backlink"><a href="#cite_ref-5">↑</a></span> <span class="reference-text">Tsai, S. Q. et al. (2014). Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nature Biotechnol. 32, 569–576 <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1038/nbt.2908">10.1038/nbt.2908</a></span></span>
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<li id="cite_note-6"><span class="mw-cite-backlink"><a href="#cite_ref-6">↑</a></span> <span class="reference-text">Guilinger, J. P., Thompson, D. B. & Liu, D. R. (2014). Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Nature Biotechnol. 32, 577–582 <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1038/nbt.2909">10.1038/nbt.2909</a></span></span>
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<li id="cite_note-7"><span class="mw-cite-backlink"><a href="#cite_ref-7">↑</a></span> <span class="reference-text">Wyvekens, N., Topkar, V. V., Khayter, C., Joung, J. K. & Tsai, S. Q. (2015). Dimeric CRISPR RNA-guided FokI-dCas9 nucleases directed by truncated gRNAs for highly specific genome editing. Hum. Gene Ther. 26, 425–431 <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1089/hum.2015.084">10.1089/hum.2015.084</a></span></span>
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