{"id":159,"date":"2018-05-03T11:23:36","date_gmt":"2018-05-03T09:23:36","guid":{"rendered":"https:\/\/site.uit.no\/cube\/?page_id=159"},"modified":"2018-05-03T13:31:42","modified_gmt":"2018-05-03T11:31:42","slug":"wp4-experimental-design","status":"publish","type":"page","link":"https:\/\/site.uit.no\/cube\/about\/wp4-experimental-design\/","title":{"rendered":"WP4 Experimental design"},"content":{"rendered":"<p>Together with NorStruct (Dr. Altermark) and ArcticZymes, the production and characterization of\u00a0enzymes in terms of catalytic efficiency and thermal stability will carried out. Mutations suggested to\u00a0be important for either catalytic activity and tuning of the enthalpy-entropy balance (<a href=\"https:\/\/site.uit.no\/cube\/about\/wp1-reaction-rates\/\">WP1<\/a>) or thermal\u00a0stability (<a href=\"https:\/\/site.uit.no\/cube\/about\/wp2-point-mutations\/\">WP2<\/a>) will be produced and assayed. Both collaborators have extensive hands-on expertise from\u00a0working with the relevant enzymes. Production protocols with good yields and kinetic assays are available for both\u00a0cold- and warm-active enzyme.<\/p>\n<p><img fetchpriority=\"high\" decoding=\"async\" class=\"wp-image-188 alignright\" src=\"https:\/\/site.uit.no\/cube\/wp-content\/uploads\/sites\/364\/2018\/05\/bfac2-1024x676.png\" alt=\"\" width=\"824\" height=\"544\" srcset=\"https:\/\/site.uit.no\/cube\/wp-content\/uploads\/sites\/364\/2018\/05\/bfac2-1024x676.png 1024w, https:\/\/site.uit.no\/cube\/wp-content\/uploads\/sites\/364\/2018\/05\/bfac2-300x198.png 300w, https:\/\/site.uit.no\/cube\/wp-content\/uploads\/sites\/364\/2018\/05\/bfac2-768x507.png 768w, https:\/\/site.uit.no\/cube\/wp-content\/uploads\/sites\/364\/2018\/05\/bfac2.png 1878w\" sizes=\"(max-width: 824px) 100vw, 824px\" \/><\/p>\n<p>With our international collaborators we will start out to\u00a0characterise changes in the translational machinery due to cold environments. So far, there is basically no\u00a0data available for the performance of the translational apparatus in psychrophilic species but\u00a0there are clear sequence characteristics found for the 16S rRNA. Thus, the project will attempt to\u00a0isolate and culture psychrophilic bacteria from, e.g., Vibrio and Psychrobacter species in order to extract\u00a0components of the translation system. With our in-house in vitro translation capabilities it will then be\u00a0possible to examine the speed of protein synthesis both for full reporter genes and short synthetic mRNAs\u00a0with fast kinetics methods. The effects of external factors such as temperature and pressure can then be\u00a0studied. \u00a0Evolutionary\u00a0experiments on E. Coli and Salmonella will also be attempted, where these are submitted to cold growth conditions and selected for\u00a0fitness in many rounds. With subsequence genome sequencing relevant genetic changes both in the\u00a0translation machinery, which is usually limiting for growth in these species, and regular enzymes can be\u00a0detected and analyzed. This will provide highly interesting data for subsequent computational analysis in\u00a0<a href=\"https:\/\/site.uit.no\/cube\/about\/wp1-reaction-rates\/\">WP1<\/a>.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Together with NorStruct (Dr. Altermark) and ArcticZymes, the production and characterization of\u00a0enzymes in terms of catalytic efficiency and thermal stability will carried out. Mutations suggested to\u00a0be important for either catalytic activity and tuning of the enthalpy-entropy balance (WP1) or thermal\u00a0stability &hellip; <a href=\"https:\/\/site.uit.no\/cube\/about\/wp4-experimental-design\/\">Continued<\/a><\/p>\n","protected":false},"author":862,"featured_media":0,"parent":26,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"page-fullwidth.php","meta":{"footnotes":""},"class_list":["post-159","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/pages\/159","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/users\/862"}],"replies":[{"embeddable":true,"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/comments?post=159"}],"version-history":[{"count":3,"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/pages\/159\/revisions"}],"predecessor-version":[{"id":189,"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/pages\/159\/revisions\/189"}],"up":[{"embeddable":true,"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/pages\/26"}],"wp:attachment":[{"href":"https:\/\/site.uit.no\/cube\/wp-json\/wp\/v2\/media?parent=159"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}