{"id":2249,"date":"2014-06-04T19:59:57","date_gmt":"2014-06-04T23:59:57","guid":{"rendered":"https:\/\/www.med.unc.edu\/csb\/pep\/services\/expression-services\/"},"modified":"2024-02-05T12:19:42","modified_gmt":"2024-02-05T17:19:42","slug":"expression-services","status":"publish","type":"page","link":"https:\/\/www.med.unc.edu\/csb\/pep\/services\/expression-services\/","title":{"rendered":"Expression Services"},"content":{"rendered":"<p>The facility currently utilizes <em>E. coli<\/em> and mammalian cell expression systems. Typically, users provide a sequence-verified expression vector containing the gene(s) of interest. Alternatively, our staff can undertake expression vector construction if requested.<\/p>\n<h2><em>E. coli <\/em>Expression<\/h2>\n<ul>\n<li>Small-scale test expressions can be conducted to determine optimal expression conditions.<\/li>\n<li>Large multi-liter shaking, suspension cultures of <em>E. coli<\/em> can be grown and processed at temperatures ranging from 18\u00b0C to 42\u00b0C. Max capacity is 18L at a time.<\/li>\n<li>The facility stocks a variety of <em>E. coli<\/em> strains to address specific issues like rare-codon usage, protein misfolding, and expression in specialized media, for example, for the production of selenomethionine variants or 13C and 15N labeling.<\/li>\n<\/ul>\n<h2>Mammalian Cell Expression<\/h2>\n<ul>\n<li>Our go-to cell lines are Expi293 (HEK293) and ExpiCHO (CHO-S). Both are mammalian cell lines selected for high protein expression and dense cell growth, and both are adapted for suspension culture.<\/li>\n<li>We can culture other suspension or adherent cell lines if special circumstances are required, such as limiting glycosylation or enabling DHFR stable cell selection.<\/li>\n<li>Small-scale test expressions can be performed with virtually any suspension or adherent culture. Typically these are done for a simple &#8216;yes\/no&#8217; answer to the question of whether a protein expresses, but they can be adapted for very small scale preps.<\/li>\n<li>For large scale expression experiments in suspension culture, we have a maximum of ~15L capacity.<\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>The facility currently utilizes E. coli and mammalian cell expression systems. Typically, users provide a sequence-verified expression vector containing the gene(s) of interest. Alternatively, our staff can undertake expression vector construction if requested. E. coli Expression Small-scale test expressions can be conducted to determine optimal expression conditions. Large multi-liter shaking, suspension cultures of E. coli &hellip; <a href=\"https:\/\/www.med.unc.edu\/csb\/pep\/services\/expression-services\/\" aria-label=\"Read more about Expression Services\">Read more<\/a><\/p>\n","protected":false},"author":22429,"featured_media":0,"parent":2238,"menu_order":2,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"footnotes":"","_links_to":"","_links_to_target":""},"class_list":["post-2249","page","type-page","status-publish","hentry","odd"],"acf":[],"_links_to":[],"_links_to_target":[],"_links":{"self":[{"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/pages\/2249","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/users\/22429"}],"replies":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/comments?post=2249"}],"version-history":[{"count":0,"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/pages\/2249\/revisions"}],"up":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/pages\/2238"}],"wp:attachment":[{"href":"https:\/\/www.med.unc.edu\/csb\/pep\/wp-json\/wp\/v2\/media?parent=2249"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}