{"id":152,"date":"2022-03-22T18:01:43","date_gmt":"2022-03-22T18:01:43","guid":{"rendered":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/?p=152"},"modified":"2022-03-22T18:21:30","modified_gmt":"2022-03-22T18:21:30","slug":"library-of-influenza-h3-pseudotypes-with-an-hiv-backbone","status":"publish","type":"post","link":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/","title":{"rendered":"Library of Influenza H3 pseudotypes with an HIV backbone"},"content":{"rendered":"<p><strong>Strain (accession, expression plasmid, protease for HA cleavage)<\/strong><\/p>\n<p>A\/Texas\/50\/2012 (KC892952.1 pI.18 TMPRSS4)<\/p>\n<p>A\/Udorn\/307\/1972 (DQ508929.1 pI.18 TMPRSS2)<\/p>\n<p>A\/California\/7\/2004 (CY114373.1 pEVAC HAT)<\/p>\n<p>A\/Wisconsin\/67\/2005 (CY034116.1 pEVAC HAT)<\/p>\n<p>A\/Japan\/WRAIR1059P\/2008 (pEVAC HAT)<\/p>\n<p>A\/Switzerland\/9715293\/2013 (EPI814528 pEVAC HAT)<\/p>\n<p>A\/New Caledonia\/71\/2014 (EPI551570 pEVAC HAT)<\/p>\n<p>A\/duck\/Quang Ninh\/220\/2014 (LC053492 pEVAC HAT)<\/p>\n<p>A\/ruddy turnstone\/Delaware Bay\/606\/2017 (MH135712 pEVAC TMPRSS2)<\/p>\n<p>A\/Kansas\/14\/2017 Vaccine strain, 3c3.A (pEVAC HAT)<\/p>\n<p>A\/Switzerland\/8060\/2017 (EPI1326015 pEVAC TMPRSS4)<\/p>\n<p>A\/South Australia\/34\/2019 (EPI1607117 pEVAC HAT)<\/p>\n<p>&nbsp;<\/p>\n<p><img src=\"https:\/\/www.mdpi.com\/vaccines\/vaccines-09-00741\/article_deploy\/html\/images\/vaccines-09-00741-g002.png\" \/><\/p>\n<p>&nbsp;<\/p>\n<p>Schematic representation of the production of influenza H3 pseudotypes by plasmid transfection. (a) Using a 4 plasmid system, exogenous NA was added to the transfected cultures at least 8 h post-transfection. (b) Addition of protease, which is necessary for the production of H3 pseudotypes is optimized to increase production titres by transfecting using a \u2018checkerboard\u2019 approach with different proteases (e.g., <strong>HAT, TMPRSS4, <\/strong>and<strong> TMPRSS2)<\/strong>. Protease plasmid was added at a ratio of 1:1, 1:0.5, and 1:0.25 to HA plasmid DNA for rapid optimization in a 6 well plate format. All pseudotypes were harvested after 48 h in culture. ORCID: https:\/\/orcid.org\/0000-0002-7978-3815<\/p>\n<ul class=\"kent-social-links\"><li><a href='http:\/\/www.facebook.com\/sharer.php?u=https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/&amp;t=Library of Influenza H3 pseudotypes with an HIV backbone' target='_blank'><i class='ksocial-facebook' title='Share via Facebook'><\/i><\/a><\/li><li><a href='http:\/\/twitter.com\/home?status=Library of Influenza H3 pseudotypes with an HIV backbone%20https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/' target='_blank'><i class='ksocial-twitter' title='Share via Twitter'><\/i><\/a><\/li><li><a href='https:\/\/plus.google.com\/share?url=https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/' target='_blank'><i class='ksocial-google-plus' title='Share via Google Plus'><\/i><\/a><\/li><li><a href='http:\/\/linkedin.com\/shareArticle?mini=true&amp;url=https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/&amp;title=Library of Influenza H3 pseudotypes with an HIV backbone' target='_blank'><i class='ksocial-linkedin' title='Share via Linked In'><\/i><\/a><\/li><li><a href='mailto:content=https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/&amp;title=Library of Influenza H3 pseudotypes with an HIV backbone' target='_blank'><i class='ksocial-email' title='Share via Email'><\/i><\/a><\/li><\/ul>","protected":false},"excerpt":{"rendered":"<p>Strain (accession, expression plasmid, protease for HA cleavage) A\/Texas\/50\/2012 (KC892952.1 pI.18 TMPRSS4) A\/Udorn\/307\/1972 (DQ508929.1 pI.18 TMPRSS2) A\/California\/7\/2004 (CY114373.1 pEVAC HAT) A\/Wisconsin\/67\/2005 (CY034116.1 pEVAC HAT) A\/Japan\/WRAIR1059P\/2008 &hellip; <a href=\"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-h3-pseudotypes-with-an-hiv-backbone\/\">Read&nbsp;more<\/a><\/p>\n","protected":false},"author":2395,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[194094,1],"tags":[],"_links":{"self":[{"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/posts\/152"}],"collection":[{"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/users\/2395"}],"replies":[{"embeddable":true,"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/comments?post=152"}],"version-history":[{"count":3,"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/posts\/152\/revisions"}],"predecessor-version":[{"id":159,"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/posts\/152\/revisions\/159"}],"wp:attachment":[{"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/media?parent=152"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/categories?post=152"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/wp-json\/wp\/v2\/tags?post=152"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}