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. 2021 Aug 3;9(8):850. doi: 10.3390/vaccines9080850

Figure 1.

Figure 1

Generation and characterization of scPR8 containing receptor binding domain (RBD) of SARS-CoV-2 spike protein (scPR8-RBD-M2). (A) Schematic diagram of a reverse genetics-generated scPR8-RBD-M2. The chimeric gene cassette comprising tPA followed by a bicistronic RBD and M2 protein expression was introduced in the segment 4 (HA). The packaging signal sequence (Ψ) at both 3′ and 5′ terminal ends are also depicted. (B). scPR8-RBD-M2 infected MDCK-HA cells were assessed for RBD-HAcyt expression by (B)IFA using α-RBD and α-spike antibodies (scale bar indicates 100 μm) and (C) Western blot using α-RBD, -2A, -NP and –β actin as primary antibodies. Lower band(s) in lane 2 detected by α-2A antibody possibly indicate partially degraded mCherry-2A protein. (D) Stability of RBD-M2 inserted in infected MDCK-HA cells after several passages and (E) at the 8th passage was determined by Western blot. Mock infected MDCK-HA cells were used as a control. WCL: whole cell lysate.