Author: Mateo, Roberto; Nagamine, Claude M.; Kirkegaard, Karla
Title: Suppression of Drug Resistance in Dengue Virus Document date: 2015_12_15
ID: 6bx2nrui_32
Snippet: Plasmids and RNA transcription. Viral cDNA manipulation and generation of infectious RNA for tissue culture experiments have been described elsewhere (30) . A silent mutation in the core protein region of the genome that introduced an AflII restriction site in the wild-type genome and all drug resistance mutations were introduced using the QuikChange site-directed mutagenesis kit (Agilent Technologies, Santa Clara, CA). For mouse experiments, ser.....
Document: Plasmids and RNA transcription. Viral cDNA manipulation and generation of infectious RNA for tissue culture experiments have been described elsewhere (30) . A silent mutation in the core protein region of the genome that introduced an AflII restriction site in the wild-type genome and all drug resistance mutations were introduced using the QuikChange site-directed mutagenesis kit (Agilent Technologies, Santa Clara, CA). For mouse experiments, serotype 2 strain PL046 was used. Envelope muta- tions N124D and K128E, which confer increased pathogenicity in dengue virus-susceptible 129 IFNAR Ϫ/Ϫ IFNGR Ϫ/Ϫ mice (31), were introduced as described above. Each amplified DNA segment was sequenced in its entirety to ensure that no adventitious mutations were introduced and was then reintroduced into the infectious cDNA backbone to generate infectious RNA, as described by Mateo et al. in 2013 (30) .
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