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J Gen Virol 74 (1993), 1371-1379; DOI 10.1099/0022-1317-74-7-1371
© 1993 Society for General Microbiology
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Stable constitutive expression of glycoprotein B (gpUL55) of human cytomegalovirus in permissive astrocytoma cells

B. Reis, E. Bogner, M. Reschke, A. Richter, Th. Mockenhaupt and K. Radsak

Institut für Virologie der Philipps-Universität, Robert-Koch-Strasse 17, 355 Marburg, Germany

Permanent cell lines showing homogeneous constitutive expression of glycoprotein B (gpUL55; gB) of human cytomegalovirus (HCMV) were selected, in the presence of geneticin, from human astrocytoma cells (U373) after transfection with recombinant pRC/CMV-gB carrying the complete coding sequence for HCMV gB and for aminoglycoside phosphotransferase. The biosynthesis and processing including specific proteolytic cleavage, formation of disulphide-linked oligomers as well as transport of recombinant gB in three of four established transformed cell lines essentially resembled that found in infected parental U373 except for eventual degradation after 2 h of gB synthesis. Analysis of the fourth transformant expressing uncleaved gB suggested that proteolytic cleavage is not required for normal intracellular transport. The stable transformants retained permissiveness for productive superinfection with HCMV. The application of cell lines transformed with mutagenized HCMV gB for the rescue of genetically engineered HCMV mutants is discussed.

Received 21 December 1992; accepted 19 February 1993.


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