Development of a cell sorting procedure to increase the sensitivity of detection of protein-protein interactions in plant protoplasts

作者：Zhang Xin; Wong Sek Man^*

来源：Journal of Virological Methods, 2011, 173(2): 347-352.

DOI：10.1016/j.jviromet.2011.03.007

• Hibiscus chlorotic ringspot virus
• Flow cytometry
• Subcellular localization
• Yellow fluorescence
• Transfection efficiency

To visualize subcellular localization of viral proteins and interactions between viral proteins and host proteins in vivo, transfection of plasmids into protoplasts to over-express transiently fusion proteins with a fluorescent tag is a common method. However, due to the low efficiency (0.1-3.0%) of plasmid transfection into protoplasts, it is difficult to identify protoplasts that emit fluorescence using confocal microscopy. A flow cytometry sorting protocol was developed for separating kenaf protoplasts that emit yellow fluorescence. The sorted protoplasts showed strong fluorescence and the protoplasts were intact This will improve the use of confocal microscopy for studying subcellular localization and protein interactions in protoplasts isolated from plants with low transfection efficiency.

• 出版日期2011-5