Figure 1

Subcellular localization of the wild-type and truncated HIV integrase fused with YFP. A) Schematic structure of HIV-1 integrase-YFP fusion proteins. Full-length (1–288aa) HIV-1 integrase, the N-terminus-truncated mutant (51–228aa) or the C-terminus-truncated mutant (1–212aa) was fused in frame at the N-terminus of YFP protein. The cDNA encoding for each IN-YFP fusion protein was inserted in a SVCMV expression plasmid. B) Expression of different IN-YFP fusion proteins in 293T cells. 293T cells were transfected with each IN-YFP expressor and at 48 hours of transfection, cells were lysed, immunoprecipitated with anti-HIV serum and resolved by electrophoresis through a 12.5% SDS-PAGE followed by Western blot with rabbit anti-GFP antibody. The molecular weight markers are indicated at the left side of the gel. C) Intracellular localization of different IN-YFP fusion proteins. HeLa cells were transfected with each HIV-1 IN-YFP fusion protein expressor and at 48 hours of transfection, cells were fixed and subjected to indirect immunofluorescence using rabbit anti-GFP and then incubated with FITC-conjugated anti-rabbit antibodies. The localization of each fusion protein was viewed by Fluorescence microscopy with a 50× oil immersion objective. Upper panel is fluorescence images and bottom panel is DAPI nucleus staining.