Author: Schwarz, Megan C.; Sourisseau, Marion; Espino, Michael M.; Gray, Essanna S.; Chambers, Matthew T.; Tortorella, Domenico; Evans, Matthew J.
Title: Rescue of the 1947 Zika Virus Prototype Strain with a Cytomegalovirus Promoter-Driven cDNA Clone Document date: 2016_9_28
ID: 0i1abjfa_6
Snippet: Identification and stabilization of a bacterially toxic ZIKV cDNA region. To generate a plasmid carrying the ZIKV cDNA (Fig. 1B) , we purified RNA from MR766 inoculum and used reverse transcription-PCR (RT-PCR) to amplify overlapping regions of the viral genome, which were progressively cloned into the high-copy-number bacterial plasmid pCDNA6.2 (Fig. 1C) . In this plasmid, the authentic 5= end of the viral sequence was placed at the transcriptio.....
Document: Identification and stabilization of a bacterially toxic ZIKV cDNA region. To generate a plasmid carrying the ZIKV cDNA (Fig. 1B) , we purified RNA from MR766 inoculum and used reverse transcription-PCR (RT-PCR) to amplify overlapping regions of the viral genome, which were progressively cloned into the high-copy-number bacterial plasmid pCDNA6.2 (Fig. 1C) . In this plasmid, the authentic 5= end of the viral sequence was placed at the transcriptional initiation site of the cytomegalovirus (CMV) promoter, the simian virus 40 (SV40) polyadenylation site terminates transcription, and an HDVr trims the viral genome to its authentic 3= end (Fig. 1B) . The resulting transcript of the ZIKV genome is identical to that of a genome deposited in a host cell by a virion and is expected to initiate infection.
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