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Expression and accumulation of heterologous molecules in the protein storage vacuoles of soybean seeds

External protocol Created on 30 Apr 2014

Authors

Giovanni Vianna, Nicolau da Cunha, and Elíbio Rech

Summary

The production of recombinant molecules in seeds plant has presented interesting results related to the synthesis, secretion, compartmentalisation, post-translational modifications and purification, scalability, stable expression, the absence of contaminants and human pathogens, and knowledge of the growth, harvest, storage and processing practices. In this context, the soybean plant has emerged as an option to express recombinant proteins. We obtained several transgenic soybean plant lines, expressing different types of recombinant molecules, under the control of the tissue-specific soybean seed storage β-conglycinin promoter. This promoter is the most abundant protein in soybean seeds, directing the expression of the molecules that accumulate in the seed storage tissue. Cotyledonary immunocytochemical analysis of the seeds demonstrated that the targeted proteins effectively drove polypeptide accumulation into the protein storage vacuoles (PSVs). Here, we provide a detailed protocol to reproduce the results described above and properly direct the expression of recombinant protein into the PSVs.

Further details

The protocol was published on Protocol Exchange on 1 February 2011. To see the entire protocol, click on the source link.

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