{"id":154,"date":"2022-03-22T18:16:43","date_gmt":"2022-03-22T18:16:43","guid":{"rendered":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/?p=154"},"modified":"2022-03-22T18:21:50","modified_gmt":"2022-03-22T18:21:50","slug":"library-of-influenza-b-pseudotypes-with-an-hiv-backbone","status":"publish","type":"post","link":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2022\/03\/22\/library-of-influenza-b-pseudotypes-with-an-hiv-backbone\/","title":{"rendered":"Library of Influenza B pseudotypes with an HIV backbone"},"content":{"rendered":"

Strain (accession, expression plasmid, protease for HA cleavage)<\/strong><\/p>\n

B\/Hong Kong\/8\/1973 (K00425 phCMV1 HAT)<\/p>\n

B\/Victoria\/2\/1987 (FJ766840 phCMV1 HAT)<\/p>\n

B\/Yamagata\/16\/1988 (CY018765.1 phCMV1 HAT)<\/p>\n

B\/Florida\/4\/2006 (EU515992 phCMV1 HAT)<\/p>\n

B\/Bangladesh\/3333\/2007 (CY115255.1 pI.18 HAT)<\/p>\n

B\/Brisbane\/60\/2008 (KX058884.1 pI.18 HAT)<\/p>\n

B\/Brisbane\/60\/2008 (EPI753679 pEVAC HAT)<\/p>\n

B\/Phuket\/3073\/2013 (pEVAC HAT)<\/p>\n

B\/Colorado\/06\/2017 (pEVAC HAT)<\/p>\n

B\/Washington\/2\/2019 (EPI1368874 pEVAC HAT)<\/p>\n

 <\/p>\n

<\/p>\n

Schematic representation of the production of influenza B 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 influenza B pseudotypes is optimized to increase production titres by transfecting using a \u2018checkerboard\u2019 approach with different proteases (e.g., HAT<\/strong>, TMPRSS4, and TMPRSS2). 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<\/a><\/p>\n