Identification of simian varicella virus gene 21 promoter region using green fluorescent protein

Clinical, pathological, immunological and virological features of simian va ricella virus (SVV) infection in primates closely resemble those of varicel la tester virus (VZV) infection in humans. In ganglia infected latently of humans and monkeys, gene 21 of VZV and SVV is transcribed, respectively. We determined the nucleotide sequence of the intragenic region between SW gen es 20 and 21 to identify the putative promoter region for SW gene 21. A rec ombinant clone was prepared in which the gene encoding green fluorescent pr otein (GFP) was inserted ten base pairs upstream of the predicted translati onal start site for SW gene 21. SW-infected monkey kidney cells transfected with the recombinant clone showed the presence of green fluorescence, wher eas transfection of these cells with a construct containing the GFP gene in the opposite orientation, produced no fluorescence. The recombinant clone containing GFP flanked by SW sequences can be used to prepare a SW mutant i n which the virus gene 21 promoter drives GFP. Such a mutant will be useful in analyzing varicella pathogenesis and latency in experimentally infected animals, studies not possible in humans. (C) 2000 Elsevier Science B.V. Al l rights reserved.