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Figure 7 | Retrovirology

Figure 7

From: Sequential emergence and clinical implications of viral mutants with K70E and K65R mutation in reverse transcriptase during prolonged tenofovir monotherapy in rhesus macaques with chronic RT-SHIV infection

Figure 7

Segregation of K65R and K70E mutations, and linkage of codon 68 mutations with K65R. Plasma viral RNA samples in which real-time PCR assays detected both K65R and K70E mutations were analyzed further; representative samples are shown. Panel A: animal 30007, week 8 of tenofovir treatment (see Figure 6). Population sequencing revealed a mixture of wild-type and mutant variants at both codons 65 and 70 (top graph); the bar indicates the codon reading frame. The selective amplification of virus sequences containing 65R or 70E by real-time PCR allowed for their enrichment from the virus background quasispecies. Direct sequencing of the mutation-specific amplicons revealed that the 65R amplicon (AGA, arginine) had wild-type sequence at codon 70 (AAA, lysine; middle graph), while the 70E amplicon (GAA, glutamic acid) had wild-type at codon 65 (lysine, AAA; bottom graph). Thus, the K65R and K70E mutations were on separate viral genomes. Panel B: animal 30478, week 12 of tenofovir treatment. The mutation-specific amplicons from this specimen also exhibited segregation of K65R and K70E. The sequence of the 65R amplicon demonstrated mutations at codon 68 (middle graph), while the 70E amplicon had wild-type sequence (AGT, serine) at codon 68 (bottom graph). The presence of mixtures is indicated (M is A or C; R is A or G).

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