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 (EGFP) was introduced into a human immunodeficiency virus type 1 (HIV-1) molecular clone by replacing the nef gene. The EGFP-expressing HIV-1 replicated efficiently in established human T cells. The expression of EGFP in the virus-infected cells was confirmed by fluorescence microscopy and also by Western blot analysis using the GFP antibodies. The EGFP gene was stably maintained in the viral genome during prolonged passages. EGFP-expressing HIV-1 could be used for anti-HIV-1 drug screening, sorting of virus-infected cells by fluorescence activated cell sorting (FACS) analysis, and for the rapid and simple detection of virus-infected cells by fluorescence microscopy.