{"id":2945,"date":"2017-03-27T14:53:30","date_gmt":"2017-03-27T13:53:30","guid":{"rendered":"http:\/\/roboblog.fatal-fury.de\/?p=2945"},"modified":"2017-03-27T15:13:54","modified_gmt":"2017-03-27T14:13:54","slug":"biologie-silver-assembly","status":"publish","type":"post","link":"http:\/\/roboblog.fatal-fury.de\/?p=2945","title":{"rendered":"Biologie++ - Silver Assembly"},"content":{"rendered":"<blockquote><p>\nIm Glas rekombinationsmthoden haben eine Einschritt Erstellung von gro\u00dfen DNA Sequenezen<br \/>\nvon mehreren einzelnen erm\u00f6glicht. Obwohl synthetische biologische Schaltungen im Prinzip<br \/>\nauf der selben Art erstellt werden k\u00f6nnen, enthalten sie normalerweise sich wiederholende Sequenzteile,<br \/>\nwie promoters und terminator, die bei der Recombination st\u00f6ren.<br \/>\nWir benutzen ein Computer gest\u00fctzten Ansatz um biologische, synthetische inaktive unique nucleotide<br \/>\nsequences (UNSes) zu designen, die das genaue Erstellen erleichtern.<br \/>\nWichtig: unsere designten UNSes machen es m\u00f6glich Teile zu assemblieren die wiederholende<br \/>\nterminator und insulator Sequenzen enthalten. Und damit isolierte, funktionale genetische Schaltungen<br \/>\nin Bakterien und S\u00e4ugetierzellen zu erstellen.<br \/>\n...\n<\/p><\/blockquote>\n<blockquote><p>\nIn vitro recombination methods have enabled one-<br \/>\nstep construction of large DNA sequences from<br \/>\nmultiple parts. Although synthetic biological<br \/>\ncircuits can in principle be assembled in the same<br \/>\nfashion, they typically contain repeated sequence<br \/>\nelements such as standard promoters and termin-<br \/>\nators that interfere with homologous recombination.<br \/>\nHere we use a computational approach to design<br \/>\nsynthetic, biologically inactive unique nucleotide<br \/>\nsequences (UNSes) that facilitate accurate ordered<br \/>\nassembly. Importantly, our designed UNSes make it<br \/>\npossible to assemble parts with repeated terminator<br \/>\nand insulator sequences, and thereby create<br \/>\ninsulated functional genetic circuits in bacteria and<br \/>\nmammalian cells. Using UNS-guided assembly to construct<br \/>\nrepeating promoter-gene-terminator parts, we systematically varied gene expression to<br \/>\noptimize production of a deoxychromoviridans bio-<br \/>\nsynthetic pathway in Escherichia coli. We then used<br \/>\nthis system to construct complex eukaryotic AND-<br \/>\nlogic gates for genomic integration into embryonic<br \/>\nstem cells. Construction was performed by using<br \/>\na standardized series of UNS-bearing BioBrick-<br \/>\ncompatible vectors, which enable modular assembly<br \/>\nand facilitate reuse of individual parts. UNS-guided<br \/>\nisothermal assembly is broadly applicable to the<br \/>\nconstruction and optimization of genetic circuits<br \/>\nand particularly those requiring tight insulation,<br \/>\nsuch as complex biosynthetic pathways, sensors,<br \/>\ncounters and logic gates.<\/p>\n<\/blockquote>\n<p>[1] Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Silver et al.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Im Glas rekombinationsmthoden haben eine Einschritt Erstellung von gro\u00dfen DNA Sequenezen von mehreren einzelnen erm\u00f6glicht. Obwohl synthetische biologische Schaltungen im Prinzip auf der selben Art erstellt werden k\u00f6nnen, enthalten sie normalerweise sich wiederholende Sequenzteile, wie promoters und terminator, die bei der Recombination st\u00f6ren. Wir benutzen ein Computer gest\u00fctzten Ansatz um biologische, synthetische inaktive unique nucleotide [&hellip;]<\/p>\n","protected":false},"author":2,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[43],"class_list":["post-2945","post","type-post","status-publish","format-standard","hentry","category-allgemein","tag-biologie"],"_links":{"self":[{"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=\/wp\/v2\/posts\/2945","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=2945"}],"version-history":[{"count":2,"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=\/wp\/v2\/posts\/2945\/revisions"}],"predecessor-version":[{"id":2947,"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=\/wp\/v2\/posts\/2945\/revisions\/2947"}],"wp:attachment":[{"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=2945"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=2945"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/roboblog.fatal-fury.de\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=2945"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}