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Updated: Jul 20, 2026

A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
Published on: August 31, 2014
Coexistence of the K65R/L74V and/or K65R/T215Y mutations on the same HIV-1 genome
Mireille Henry1, Christian Tourres, Philippe Colson
1Laboratory of Virology, Timone Hospital and CNRS UMR 6020, 264 Rue St Pierre, 13385 Marseille Cedex 05, France.
Objective:
To determine to which extent the mutations K65R, L74V/I and T215Y/F are linked to the same HIV-1 genome.
Methods:
Retrospective analysis of the Marseille database (8866 sequences from 3720 patients). The HIV-1 pol gene from four patients' viruses harbouring the double mutant (pure species or genotypic mixtures) was amplified, cloned and sequenced.
Results:
The four patients had a mean viral load of 5.6 log(10)copies/ml. Analysis of 73 clones (patient 1: 12 clones; patient 2: 27 clones; patient 3: 13 clones; patient 4: 21 clones) showed that 29 clones harboured K65R and L74V/I mutations. Twenty-three per cent of clones from the two bulk sequences harbouring K65K/R and T215T/Y genotypic mixtures contained K65R and T215Y on the same viral genome.
Conclusions:
The co-linearity of 65R and 74V or 65R and 215Y amino-acids on the same genome is rare. A high viral load (6.19 log(10)) associated with the coexistence of 65R and 74V on the same HIV-1 genome suggests possible compensatory mechanisms.
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