FIG. 7.
Insertion of HIV-1 gag upstream of the codon optimized gene affects cytoplasmic RNA levels. Cytoplasmic RNA was extracted 48 h after transfection of 293T cells (5 μg of each pSYN plasmid was used, and 1 μg of pCMV-Rev was cotransfected in some cases). The probe that was used was designed to be complementary to nt 1510 to 2290 of the codon-optimized gene (I). A probe specific for human ubiquitin was used as an internal control (II). (A) Lanes: 1, pSYNGP; 2, pSYN9; 3, pSYN10; 4, pSYN10 plus pCMV-Rev; 5, pSYN11; 6, pSYN11 plus pCMV-Rev; 7, pCMV-Rev. (B) Lanes: 1, pSYNGP; 2, pSYNGP-RRE; 3, pSYNGP-RRE plus pCMV-Rev; 4, pSYN12; 5, pSYN14; 6, pSYN14 plus pCMV-Rev; 7, pSYN13; 8, pSYN15; 9, pSYN17; 10, pGP-RRE3; 11, pSYN6; 12, pSYN9; 13, pCMV-Rev.