Plasmids expressing wild-type EBOV GP were obtained by inserting GP complementary DNA (cDNA) from the corresponding EBOV subtype (Zaire, strain Mayinga 1976, GenBank accession number U23187.1; Sudan, strain Gulu, GenBank accession number YP_138523.1; Bundibugyo, strain Uganda 2007, GenBank accession number ACI28624.1; Ivory Coast, strain Cote d’Ivoire 1994, GenBank accession number ACI28632.1; and Reston, strain Siena 1992, GenBank accession number AAC54891.1) between the Kpn I and Xba I sites of the PCMV3 mammalian expression vector (Sino Biological Inc., Beijing, China). Plasmid expressing MARV envelope protein GP was obtained by inserting codon-optimized Make Victoria MARV GP cDNA (strain Musoke, GenBank accession number YP_001531156.1) between the Hind III and Xba I sites of the PCMV3 mammalian expression vector. Plasmids expressing the wild-type HIV-1 envelope protein were obtained by inserting HIV-1 gp160 cDNA (strain NDK) between the Kpn I and Xba I sites of the PCMV3 mammalian expression vector. The HA and NA genes were amplified from IAV strain A/WSN/33(H1N1) cDNA by polymerase chain reaction and then inserted between the Kpn I and Eco RI or Xho I sites, respectively, of the pcDNA4/TO vector (Invitrogen, Carlsbad, CA). Other plasmids included pCMV-VSVG (Addgene), pNL 4-3 (NIH AIDS Reagent Program), and pAdvantage (Promega). The EBOV GP and influenza virus HA mutants were constructed with the Site-Directed Mutagenesis Kit (Agilent Technologies) and confirmed by sequencing (BGI, Beijing). All plasmids used for transfection were amplified using the Maxiprep Kit (Promega) according to the manufacturer’s instructions.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.