The study made use of previously described cultured Boa constrictor kidney cells, I/1Ki, [34 (link)] and persistently SwSCV-1- infected I/1Ki cells, I/1Ki-Δ [17 (link)]. The cells were maintained in Minimal Essential Medium Eagle (Sigma-Aldrich, St.Louis, MO, USA) supplemented with 10% fetal bovine serum (ThermoFisher Scientific, Waltham, MA, USA), 200 mM L-glutamine (Sigma-Aldrich, St. Louis, MO, USA), 100 µg/mL of streptomycin (Sigma-Aldrich, St. Louis, MO, USA), and 100 U/mL of penicillin (Sigma-Aldrich, St. Louis, MO, USA) in an incubator at 30 °C with 5% CO2.
To establish another persistently SwSCV-1-infected I/1Ki cell line, designated I/1Ki-1.2×Δ, we transfected I/1Ki cells with a plasmid containing 1.2 copies of the SwSCV-1 genome (described below) and maintained the cells as described above and earlier [17 (link)].
To study infectious particle formation of the SwSCV-1-infected cell lines, we conducted superinfection studies with HISV-1 [35 (link)], earlier demonstrated to be an efficient helper virus for SwSCV-1 [17 (link)]. The superinfection studies and detection followed the protocol described [17 (link)]. For the infection, we used 600 copies of HISV-1 S segment RNA per cell, which corresponds roughly to a multiplicity of infection (MOI) of 10.
To establish another persistently SwSCV-1-infected I/1Ki cell line, designated I/1Ki-1.2×Δ, we transfected I/1Ki cells with a plasmid containing 1.2 copies of the SwSCV-1 genome (described below) and maintained the cells as described above and earlier [17 (link)].
To study infectious particle formation of the SwSCV-1-infected cell lines, we conducted superinfection studies with HISV-1 [35 (link)], earlier demonstrated to be an efficient helper virus for SwSCV-1 [17 (link)]. The superinfection studies and detection followed the protocol described [17 (link)]. For the infection, we used 600 copies of HISV-1 S segment RNA per cell, which corresponds roughly to a multiplicity of infection (MOI) of 10.
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