Structural genomics groups have identified the need to generate multiple truncated versions of each target to improve their success in producing a well-expressed, soluble, and stable protein and one that crystallizes and diffracts to a sufficient resolution for structural determination. At the SGC, we opted for the Ligation-Independent Cloning (LIC) method which provides the medium throughput we desire to produce and screen many proteins in a parallel process. Here, we describe our LIC protocol for generating constructs in a 96-well format and provide a choice of vectors suitable for expressing proteins in both E. coli and the baculovirus expression vector system (BEVS). © 2014 Springer Science+Business Media, LLC.
CITATION STYLE
Strain-Damerell, C., Mahajan, P., Gileadi, O., & Burgess-Brown, N. A. (2014). Medium-throughput production of recombinant human proteins: Ligation-independent cloning. Methods in Molecular Biology, 1091, 55–72. https://doi.org/10.1007/978-1-62703-691-7_4
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