Computational protocol: A Chimeric Affinity Tag for Efficient Expression and Chromatographic Purification of Heterologous Proteins from Plants

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Protocol publication

[…] Structural models were generated in silico for SlCYS8 (GenBank accession no. AF198390) and tentative Cysta-tag hybrids to predict the impact of inserting a (His)6 [or 6x His] hexapeptide motif in the original cystatin structure. Twenty possible models were built for each possible variant using Modeller v. 9.7 (), with the NMR solution structure coordinates of oryzacystatin I () as a template (Protein Data Bank accession no. 1EQK). The stereochemical quality of each model was assessed by comparison with the oryzacystatin structure using the PROCHECK program, v.3.5.4 (http://www.ebi.ac.uk/thornton-srv/ software/PROCHECK/; ). The best model for SlCYS8 and the best model for a tentative Cysta-tag variant were selected for visualization purposes, Cysta-tag engineering and heterologous expression in Escherichia coli or N. benthamiana. […]

Pipeline specifications

Software tools MODELLER, PROCHECK
Application Protein structure analysis
Chemicals Cysteine