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Transient Expression of Virus‐Specific Promoters in Murine Resident Peritoneal Macrophages
Author(s) -
Wu Linxian,
Anaraki Farvardin,
Morahan Page S.,
Leary Kathryn
Publication year - 1990
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.48.3.229
Subject(s) - biology , transfection , vero cell , microbiology and biotechnology , herpes simplex virus , virus , virology , promoter , plasmid , cell culture , dna , gene expression , gene , biochemistry , genetics
Monocyte‐macrophages (MO), being non‐permissive for most viruses, play an important role in resistance to virus infection. In order to establish the mechanism of abortive infection of murine resident peritoneal MO (ResPMO) by herpes simplex virus type 1 (HSV‐1), it is desirable to transfect these cells with viral promoters linked to an assayable gene, for example, the bacterial chloramphenicol acetyl transferase (CAT) gene. This will facilitate studies designed to measure levels of promoter activation or repression in these specialized cells. Transient expression of CAT in ResPMO was achieved with DEAE‐dextran, but not using either calcium phosphate precipitate or lipofectin. CAT expression driven by various virus‐specific promoters was less efficient in ResPMO compared with Vero cells and approximately 50% of input plasmid DNA remained in Vero cells at 48 h post transfection, but only 9% was detectable in ResPMO. However, approximately 6% of ResPMO and 9% of Vero cells contained CAT‐specific DNA at 24 h post transfection. In addition, 2% of cells of either cell type contained CAT‐specific polypeptide at 48 h. This is therefore the first report that the non‐replicating murine ResPMO can be transfected in vitro and more importantly, that these cells express the transfected gene products.