Journal: Biotechnology Letters

Article Title: Production of the sheep pox virus structural protein SPPV117 in tobacco chloroplasts

doi: 10.1007/s10529-021-03117-x

Figure Lengend Snippet: Analysis of SPPV117 protein accumulation in leaves from P117 and R117 plants. a Western blot analysis of total soluble proteins from transplastomic (lanes 1–6) with SPPV117-specific antibodies. Different amounts of the bacterially produced protein were used for protein quantitation. Plant protein samples were loaded at 5 µg per lane. b Western blot using penta-his antibodies to detect SPPV117 in Ni–NTA fractions from R117 plants. L lysate, F flow throw fraction, W wash fraction, E 1 – E 5 , eluted fractions. c , d Coomassie blue-stained SDS-PAGE and Western blot using SPPV117-specific antibodies after first (lane 1) and second (lane 2) rounds of chromatography. e Western blot of purified chloroplast-produced SPPV117 protein (0.5 µg, lane 1) with serum from a sheep infected with sheep pox virus. Molecular mass marker (M) sizes are indicated in kDa. Protein samples (5 µg) from untransformed plants (wt) contained in the same buffer as proteins from transplastomic lines were used as negative control

Article Snippet: For purified proteins, murine penta-his antibodies and anti-mouse IgG conjugated with horseradish peroxidase (5-Prime, Hamburg, Germany) were used as primary and secondary antibodies, respectively at a 1:6000 dilution.

Techniques: Western Blot, Produced, Protein Quantitation, Staining, SDS Page, Chromatography, Purification, Infection, Marker, Negative Control