Production of a His-tagged canecystatin in transgenic sugarcane.

Flavio Henrique-Silva, Andrea Soares-Costa

Laboratory of Molecular Biology, Department of Genetic and Evolution, Federal University of São Carlos, São Carlos, SP, Brazil.

Methods in molecular biology (Clifton, N.J.) 2012

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Transgenic plants have been widely used as expression systems of recombinant proteins in recent years because it can be an efficient alternative for the large-scale production of proteins. This is an area with great potential but is still not much explored. Indeed, this system can bring a breakthrough in the expression of any protein. The model used here as a protein factory was sugarcane, a crop of great global importance. This chapter describes the system that has been adopted in the routine production of transgenic sugarcane coupled with protein purification protocol. In this chapter, we describe production of transgenic sugarcane expressing a His-tagged cystatin under the control of the maize ubiquitin promoter. A transformed sugarcane plant presented high levels of protein expression and was selected for the purification of this protein through affinity chromatography in a nickel column. These studies demonstrate that sugarcane can be a viable expression system for recombinant protein production and that the His-tag purification strategy used to isolate the purified protein was effective.

Citation

Flavio Henrique-Silva, Andrea Soares-Costa. Production of a His-tagged canecystatin in transgenic sugarcane. Methods in molecular biology (Clifton, N.J.). 2012;847:437-50