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Figure 3 | Genetic Vaccines and Therapy

Figure 3

From: Streamlined design of a self-inactivating feline immunodeficiency virus vector for transducing ex vivodendritic cells and T lymphocytes

Figure 3

Safety evaluation of packaging and vector constructs. A) pΔenv1 transportation from transfected to transduced cells as evaluated by gag p25 PCR using the indicated primers. Lane A: DNA from transfected 293T cells; lane B: no template control; lanes C and D: DNA and RNA from transduced 293T cells; lane E: DNA from mock transfected cells. B) Translocation of the U3 deletion to the 5'LTR, as checked by PCR using primers annealing to the beginning U3 and within the R region of LA34 proviral DNA. Lane A: pΔ00 plasmid (full-length LTR), lane B: no template control; lane C: DNA from transduced 293T cells, lane D: DNA from mock transduced cells. C) Analysis of LTR directed transcription as tested by PCR using primers upstream the GFP promoter. Lane A: DNA of transduced 293T cells; lane B: no template control; lanes C and D: RNA from transduced 293T cells with and without DNase treatment prior to reverse transcription; lanes E and F: RNA from mock transduced cells treated as for C and D. Primer nt position referred to the NC_001482 sequence. M1: 100 base-pair ladder, M2: Gene ruler 1 Kb DNA ladder (GE Healthcare).

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Genetic Vaccines and Therapy

ISSN: 1479-0556