Protein extraction from Nicotiana benthamiana leaf - Bertin Instruments

Environment

Protein extraction from Nicotiana benthamiana leaf

Sources: Institute of Genetics, Heinrich-Heine University Düsseldorf, Germany

Context

The laboratory is focused on developmental biology of plants using the model system Arabidopsis thaliana. The aim of this study was to investigate the intracellular localization of protein receptors (CLV1, CLV2, and CRN) in plant cells and their tendencies for protein-protein interactions. To analyze receptor localization and interaction, a transient expression system in Nicotiana benthamiana leaf epidermis cells was used [1].

[1] A. Bleckmann et al., Plant Physiology, January loading control. wt, wild type. 2010, Vol. 152, pp. 166–176, www.plantphysiol.org

Results

Our transient expression studies of translational fusions with GFP or mCherry now showed that all three receptor proteins can localize to the plasma membrane and have the capacity to undergo multiple interactions. The Precellys®24 is a fast method for protein isolation. Protein extracts were used to demonstrate fusion protein expression and stability.

nicotiana leaf

Figure 1: Western-blot analysis of protein extracts from N. benthamiana leaf cells transiently expressing CLV1-GFP, CLV2-GFP, or CRN-GFP. An anti-GFP antibody was used for detection; sizes of protein markers are given in kD. The Ponceau Sstained protein bands of Rubisco are shown as a loading control. wt, wild type

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