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Fig. 5 | Journal of Biological Engineering

Fig. 5

From: Heterologous expression of the isopimaric acid pathway in Nicotiana benthamiana and the effect of N-terminal modifications of the involved cytochrome P450 enzyme

Fig. 5

Analysis of protein expression in the transiently transformed leaves. Shown are SDS-PAGE (a) and immunodetection (b) of total protein extracts from crude leaf of infiltrated plants. Protein amounts corresponding to the total chlorophyll amount of 5 μg (P19, DiTPS-ISO, CYP720B4, and Barnes-CYP720B4), 10 μg (ΔTm-CYP720B4), 15 μg (Tp-Barnes-CYP720B4) and 3 μg (Tp-ΔTm-CYP720B4) were electrophoresed using SDS-PAGE. For immunodetection, Anti-FLAG antibody was used. The expected molecular masses of the FLAG-tag proteins DiTPS-ISO, CYP720B4, Barnes-CYP720B4 and ΔTm-CYP720B4 without the chloroplast transit peptide are 92, 55, 54 and 51 kDa respectively, and the masses of Tp-Barnes-CYP720B4 and Tp- ΔTm -CYP720B4 are 60 and 57 kDa respectively. The black arrow points to the visible protein band of DiTPS-ISO in SDS-PAGE. Red arrow points to faint immunodetection signal of Tp-Barnes-CYP720B4 that has presumably been processed upon transport into the chloroplasts

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Journal of Biological Engineering

ISSN: 1754-1611