|
Vector
|
IU recovered/IU addeda
|
Frequency of SIN 5' LTRd
|
Frequency of complete 5' LTRe
|
|---|
| |
Round 1b
|
Round 2c
| | |
|---|
|
pHIV-1SDmSneo
|
7.79 ± 0.62 × 10-5
|
0.21 ± 0.09
|
0% (0/17)
|
100% (17/17)
|
|
pHIV1SDmSneoLTR
|
0.71 ± 0.12
|
0.22 ± 0.40
|
0% (0/15)
|
100% (15/15)
|
|
pRK1neo
|
1.14 ± 0.47 × 10-4
|
0.11 ± 0.02
|
18% (4/22)
|
59% (13/22)
|
|
pRK1Δatt340/184neo
|
5.56 ± 1.33 × 10-5
|
2.52 ± 1.73 × 10-2
|
32% (4/11)
|
17% (2/11)
|
- aEach vector was used to transfect 293T cells and subsequently rescued by transfection with pCMVΔRnr and pHCMV-g. The virus rescued in round 1 was used in a second round of transfection/rescue. Mean ± SD. bThe titre of rescued virus as determined on A549 cells and normalised to the amount of virus initially added. cThe titre of the virus rescued in the second transfection and normalised to the amount of virus rescued in round 1. dIndividual neomycin resistant colonies from the second round titre assay were analyzed via PCR for the structure of the 5'LTR (see also Figure 8), pRK1Δatt340/184neo significantly different to pHIV-1SDmSneo (Fisher's exact Chi squared, p < 0.002). eRepair refers only to LTRs having a complete U3 sequence as determined from PCR analysis (pRK1Δatt340/184neo significantly different to pHIV1SDmSneo (p < 0.0001) and pRK1neo p < 0.05, Fisher's exact Chi squared). For pRK1neo 5 colonies produced PCR products of intermediate size and one colony out of 23 did not yield a PCR product. For pRK1Δatt340/184neo 5 colonies produced PCR products of intermediate size (see also Figure 9) and 8/20 colonies did not yield a PCR product. Colonies not producing PCR products were not scored in the results shown above.
