Flow cytometric quantification of HIV-1 Tat protein in tat-transfected Jurkat T cell lines.
The transactivator Tat protein represents a pivotal factor for the replication of human immunodeficiency virus type 1 (HIV-1). In this report, we describe a flow cytometry procedure designed to quantify the intracellular content of Tat protein in Jurkat CD4+ T lymphoblastoid cell lines, stably transfected with plasmids expressing full-length Tat protein. Various expression vectors were compared for their effectiveness to yield Tat protein in Jurkat cells, and several technical parameters were analyzed to optimize the assay. This method offers a quick and efficient approach to select stably transfected cell lines expressing different levels of specific protein.
Gibellini
D
D
Department of Clinical and Experimental Medicine, University of Bologna, Italy.
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M C
MC
Panaya
R
R
Venturi
E
E
Milani
D
D
La Placa
M
M
Zauli
G
G
D005434:Flow Cytometry / Q000379:methods; D015696:Gene Products, tat / Q000032:analysis* / Q000235:genetics; D015756:Genes, tat; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells / Q000737:chemistry* / Q000502:physiology / Q000821:virology; D010957:Plasmids; D014162:Transfection; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D014764:Viral Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus