Protocols for gene expression and protein localization in Arabidopsis. Includes: Detection of the native protein; Detection of a recombinant version; Immunofluorescence detection in Arabidopsis protoplasts; Isolation of Arabidopsis seedling protoplasts; Subcellular localization of GUS-fusion proteins in Arabidopsis seedlings; Localization of Arabidopsis proteins with GUS in situ enzyme assay. - [Read Gene Expression and Protein Localization in Arabidopsis Protocols]
Cell fractionation protocol to yield intact plant protoplasts. Technique used to purify protoplasts from the grass Glyceria fluitans. Protocol includes: Determination of leaf osmolality; Sterilization. - [Read Plant Cell Purification of Intact Plant Protoplasts]
Protocol for the transformation of Aspergillus niger. This procedure is done by first digesting the outer cell wall, forming protoplasts, and then by making holes in the membrane
through which the dna can enter using calcium chloride and polyethylene glycol. Includes: Protocol for making A.niger protoplasts; Transformation; Plating. - [Read Transformation of Aspergillus niger Protocol]
The study of transient gene expression provides a useful complement to the study of stably transformed plants. Transient assays offer a quick method of testing the effects of genes, using either phenotypic, molecular, or biochemical readouts. Transient assays based on Agrobacterium-mediated transformation of leaf explants have been described for other plant species, but it is not known how well these assays work in Arabidopsis. - [Read Transient Expression in Protoplasts]