Constructing HCV Genotype 1 Pseudoparticles

Plasma samples obtained from HCV infected subjects in the BBAASH cohort [15 (link),16 (link),45 ], Irish Anti-D cohort [46 (link)], and Swan Project [47 (link)] were used to construct a library of genotype 1 E1E2-expressing lentiviral pseudoparticles using a high-throughput production and screening approach. The E1E2 region was PCR amplified from cDNA reverse transcribed from viral RNA purified from subject plasma and cloned into the expression vector pcDNA3.2/V5/Dest (Invitrogen) using Gateway technology in a one-tube BP/LR reaction, as previously described [19 ].

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El-Diwany R., Cohen V.J., Mankowski M.C., Wasilewski L.N., Brady J.K., Snider A.E., Osburn W.O., Murrell B., Ray S.C, & Bailey J.R. (2017). Extra-epitopic hepatitis C virus polymorphisms confer resistance to broadly neutralizing antibodies by modulating binding to scavenger receptor B1. PLoS Pathogens, 13(2), e1006235.

Publication 2017

pcDNA3.2/V5/Dest

Anti d Cdna Genotype Library Plasma Vector Viral rna

Corresponding Organization : University of California, San Diego

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Variable analysis

independent variables

Plasma samples obtained from HCV infected subjects in the BBAASH cohort [15, 16, 45]
Plasma samples obtained from the Irish Anti-D cohort [46]
Plasma samples obtained from the Swan Project [47]

dependent variables

Library of genotype 1 E1E2-expressing lentiviral pseudoparticles

control variables

The E1E2 region was PCR amplified from cDNA reverse transcribed from viral RNA purified from subject plasma and cloned into the expression vector pcDNA3.2/V5/Dest (Invitrogen) using Gateway technology in a one-tube BP/LR reaction, as previously described [19]

controls

No positive or negative controls were explicitly mentioned in the provided information.

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