MOESM1 of Infection of rhesus macaques with a pool of simian immunodeficiency virus with the envelope genes from acute HIV-1 infections
Year: 2016
Type: article
Abstract: Additional file 1: Figure S1. Schematic of production of pREC_nfl_SHIVKB9Δenv/URA3. Stepwise cloning scheme to introduce the near full length SHIVKB9 genome into the pREC_nfl_HIV-1NL4-3 shuttle vector. Step 1 involves amplification of the URA3 gene from pRS316 for insertion into a digested pREC_nfl_HIV-1NL4-3 to generate pREC_nfl_HIV-1Δ5′HIV/URA3 via yeast homologous recombination. The latter vector was then digested with SacII. A PCR product containing the first half of SHIVKB9 from the primer binding site to the middle of the pol gene was inserted (step 2). URA3 fragment was inserted into pREC_nfl_5′SHIVKB9/3′HIV-1NL4-3 digested with NheI to generate pREC_nfl_5′SHIVKB9_Δ3′HIV-1/URA3 (step 3). The second half of SHIVKB9 was subsequently PCR-amplified and inserted to form pREC_nfl_SHIVKB9 vector (step 4). Finally, URA3 fragment was used to replace env gene in the produced pREC_nfl_SHIVKB9 to obtain pREC_nfl_SHIVKB9_Δenv/URA3 vector for creation of various chimeric SHIVenv viruses.
Source: Figshare
Language: English
Topic: HIV Research and Treatment
Subfield: Virology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7138369237