Team:Alberta/References/Publications/A family of LIC vectors for high through-put cloning and purification of proteins

From 2009.igem.org

William H. Eschenfeldt, Lucy Stols, Cynthia Sanville Millard, Andrzej Joachimiak and Mark I. Donnelly

from: Sharon A. Doyle (ed.), Methods in Molecular Biology: High Throughput Protein Expression and Purification, vol. 498 © 2009 Humana Press, a part of Springer Science + Business Media, Totowa, NJ

Abstract: Fifteen related ligation-independent cloning vectors were constructed for high-throughput cloning and purification of proteins. The vectors encode a TEV protease site for removal of tags that facilitate protein purification (his-tag) or improve solubility (MBP, GST). Specialized vectors allow coexpression and copurification of interacting proteins, or in vivo removal of MBP by TVMV protease to improve screening and purification. All target genes and vectors are processed by the same protocols, which we describe here.


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