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Figure 2 | Virology Journal

Figure 2

From: Nucleocapsid formation and RNA synthesis of Marburg virus is dependent on two coiled coil motifs in the nucleoprotein

Figure 2

The coiled coil region in NP is sufficient to mediate interaction with NP. (A) Schematic presentation of the constructed mutants. The coiled coil region (C1C2, grey boxes) was fused to an unrelated reporter protein (MFlag, striped box). (B) The mutants were expressed using the MVA-T7 system in HeLa cells and metabolically labeled using [35S]ProMix. Cells were lysed at 24 h post transfection and cell lysates precipitated using mouse monoclonal anti-Flag (M2) and/or mouse monoclonal anti NP (2B10) as indicated. Precipitates were separated by SDS-PAGE and analyzed using BioImager. (C) Epitope of the mouse monoclonal antibody 2B10 on NP. Mutants of NP with sequential deletions of 10 aa were expressed in HUHT7 cells and cells were lysed at 24 h post transfection. Cell lysates were separated by SDS-PAGE and gels blotted onto polyvinylidene fluoride membrane. Membrane was subjected to Western Blot analysis using either the anti MARV NP mouse monoclonal antibody 2B10 (2B10) or a rabbit anti MARV nucleocapsid antiserum (α-NC).

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