Table 4.
Summary of error rates for RNA-dependent DNA synthesis catalyzed by HIV-1 RTs, and obtained with RNAs synthesized by the T7 RNAP under different assay conditions.
| Mutation type | HIV-1BH10 RT | O_K65R/V75I RT | ||||||
|---|---|---|---|---|---|---|---|---|
| Recombinant T7 RNAP (pH 7.9; 6 mM MgCl2) | Recombinant T7 RNAP (pH 6.75; 1.5 mM MgCl2) | Recombinant T7 RNAP (pH 6.75; 1.5 mM MgCl2) | Commercial T7 RNAP a (pH 6.75; 1.5 mM MgCl2) | |||||
| No. of errors | Error rate | No. of errors | Error rate | No. of errors | Error rate | No. of errors | Error rate | |
| All classes | 47 | 1/11872 | 74 b | 1/18204 | 59 | 1/29969 | 53 | 1/44074 |
| Base substitutions | 26 | 1/13199 | 42 | 1/19726 | 40 | 1/27186 | 32 | 1/44893 |
| Transitions | 14 (54%) | 25 (60%) | 30 (75%) | 19 (59%) | ||||
| Transversions | 12 (46%) | 17 (40%) | 10 (25%) | 13 (41%) | ||||
| Frameshifts | 21 | 1/23649 | 32 | 1/37467 | 19 | 1/82826 | 21 | 1/98999 |
| Insertions | 10 (48%) | 19 (59%) | 16 (84%) | 9 (43%) | ||||
| Deletions | 11 (52%) | 13 (41%) | 3 (16%) | 12 (57%) | ||||
| At runs c | 10 (48%) | 1/10602 | 21 (66%) | 1/12188 | 15 (79%) | 1/22397 | 11 (52%) | 1/40348 |
| At non-runs | 11 (52%) | 1/35509 | 11 (34%) | 1/85726 | 4 (21%) | 1/309435 | 10 (48%) | 1/163514 |
aThe T7 RNAP used in these assays was obtained from Promega, and buffers for the corresponding RNA synthesis reactions were prepared with PIPES.
bRecombinant mutants are excluded from the analysis.
cA run is considered when there is a row of three or more identical nucleotides.