Open AccessConstruction and validation of a plasmid for HIV-1 phenotyping assays using Luminescence
Author(s) -
Bianca Cristina Duarte Vivarini,
Áislan de Carvalho Vivarini
Publication year - 2021
Publication title -
revista eletrônica acervo saúde
Language(s) - English
Resource type - Journals
ISSN - 2178-2091
DOI - 10.25248/reas.e7209.2021
Subject(s) - integrases , integrase , recombinant dna , plasmid , virology , biology , vector (molecular biology) , gene , luciferase , transfection , reporter gene , microbiology and biotechnology , computational biology , genetics , human immunodeficiency virus (hiv) , gene expression
Objective: The objective of these studies was the construction of a recombination vector for HIV-1 Integrase containing luciferase reporter gene, for the generation of recombinant viruses that can be used in phenotyping assays with integrase inhibitors. Methods: In this work, the vector pNL4-3Luc was molecularly manipulated in order to delete the integrase gene and build a vector for recombination of integrases from different virus subtypes. Results: As a result, viruses with recombinant integrases were generated in HEK293T cells transfected with plasmids, showing significant levels of luminescence. After the purification of the produced viral particles, susceptible lymphocytes were infected with the viruses containing the recombinant integrases and luminescence was detected in both integrases from HIV-1 subtype B and subtype C. Conclusion: The observation of light emission from cells infected by viruses with different integrases can be an efficient method to assess the susceptibility of these viruses in the presence of specific inhibitors for HIV-1 Integrase.