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Figure 3 | Plant Methods

Figure 3

From: A Narcissus mosaic viral vector system for protein expression and flavonoid production

Figure 3

Expression of His-tagged GUS protein (69.4 kDa) in the leaves of N. benthamiana with NMV viral vector pNMV-hGUS. A: GUS protein was extracted and separated on a 4-20% gradient SDS-PAGE gel and transferred onto PVDF membrane and was probed with Anti-β-Glucuronidase Rabbit IgG as the primary antibody; the insert wells showing blue color indicate the strength of GUS activity in each fraction. B: The identical PAGE gel as in A was used and probed with mouse monoclonal His-Tag antibody. C: GUS protein purified with Ni-Affinity Sepharose, separated on 4–20% SDS-PAGE gel was visualized with Coomassie Blue; D: An identical gel was used for the Western analysis and probed with mouse monoclonal His-Tag antibody as the primary antibody. For C and D, Lane1 Protein Marker; Lane 2: Mock Leaves; Lane 3, total protein from the leaves inoculated with pNMV-hGUS; Lane 4, the first elution of GUS from the Ni-affinity Sepharose; Lane 5–10, fractions of the GUS elution from the Ni-affinity Sepharose.

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