ZIKV virus isolate Brazil-ZKV2015 (accession number KU497555) was used to design NS1 transgenes, which were produced synthetically and codon optimized for enhanced mammalian expression by GeneArt (Germany). ZIKV NS1 was cloned into pVax (Invitrogen) downstream of the CMV promoter, and a Kozak translation initiation sequence was included. Three plasmids were generated encoding either wild-type NS1 viz. (i) pVAX-NS1, or secreted NS1 generated by the upstream introduction of the human TPA leader sequence, viz. (ii) pVAX-tpaNS1 (63). To generate secreted NS1 fused to the oligomerization domain of the C4b-p (pVAX-tpaNS1-IMX313), TPA and the oligomerization domain of C4b-p (IMX313) were introduced at the N and C termini, respectively, of the ZIKV NS1 gene as we described (35, 64). Plasmids were produced with QIAGEN endotoxin-free gigaprep kits. Sequences were confirmed by double-stranded sequencing.

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.