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J Immunol Methods. 1998 Nov 1;220(1-2):115-21.

Green fluorescent protein expressed by a recombinant vaccinia virus permits early detection of infected cells by flow cytometry.

Author information

1
Departamento de Sanidad Animal, Centro de Investigación en Sanidad Animal-INIA, Madrid, Spain.

Abstract

We have tested Green Fluorescent Protein (GFP) expressed by a vaccinia virus recombinant as a marker for viral infection. Virus recombinants expressing either wild-type GFP, or a Ser65 to Thr mutated version (GFP-S65T) were used to infect cultured cells, and the appearance of fluorescence was followed during infection by flow cytometry. Although both versions were detectable in infected cells, GFP-S65T gave up to 26-fold brighter fluorescence than wild-type GFP when excited by an argon laser beam (488 nm). In addition, GFP-S65T fluorescence appeared earlier, and infected cells could be detected above background as soon as 1 h after infection. We have used this construct to infect porcine peripheral blood lymphocytes, and show its usefulness to study virus tropism when used in combination with cell-type specific markers. Thus, GFP provides a direct, fast and convenient way to monitor infection by flow cytometry.

PMID:
9839932
DOI:
10.1016/s0022-1759(98)00156-2
[Indexed for MEDLINE]

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