FIG. 1.

Construction and expression of proviral clone HIV Gag-iGFP. (A) GFP is inserted into HIV-1(NL4-3) between the MA and CA domains of Gag with protease cleavage sites (SQNYPIVQ) created at both ends of GFP. (B) Viral production from HIV Gag-iGFP or native HIV-1 was determined by p24 ELISA of supernatants from provirus-transfected 293T cells. (C) Cell lysates from HIV- or HIV Gag-iGFP-transfected 293T cells were examined by Western blotting with anti-HIV patient serum (left) or anti-GFP antibody (right). (D) Virus particles purified through a 20% sucrose cushion were analyzed by Western blotting with anti-HIV serum (α-HIV) (left) and anti-GFP (α-GFP) (right). Viral supernatants and cell lysates were collected at 48 hpt. LTR, long terminal repeat.