GENERATION OF THE REPLICATION-COMPETENT HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 WHICH EXPRESSES A JELLYFISH GREEN FLUORESCENT PROTEIN

The gene encoding a red-shifted green fluorescent protein variant (EGF P) was introduced into a human immunodeficiency virus type 1 (HIV-1) m olecular clone by replacing the nef gene. The EGFP-expressing HIV-1 re plicated efficiently in established human T cells. The expression of E GFP in the virus-infected cells was confirmed by fluorescence microsco py and also by Western blot analysis using the GFP antibodies. The EGF P gene was stably maintained in the viral genome during prolonged pass ages. EGFP-expressing HIV-1 could be used for anti-HlV-1 drug screenin g, sorting of virus-infected cells by fluorescence activated cell sort ing (FACS) analysis, and for the rapid and simple detection of virus-i nfected cells by fluorescence microscopy. (C) 1997 Academic Press.