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Protein expression, purification & charcterisation
Accessing large quantities of pure proteins can be critical for biopharmaceutical programmes and as key components of high-throughput screens. Choice of expression system & purification strategy can
dramatically improve your chance of success. CBV has experience of developing in-house skills & expertise & in out-sourcing projects to facilitate project delivery.
Expression of a protein that regulates gene expression
The protein is "toxic" & cannot be overexpressed
Use an expression vector with tight regulation, a brief & partial induction regimen with optimised media to maximise expression yield, coupled to development of a rapid & efficient purification system allowing gramme quantities of protein to be purified for biochemical & structural studies
Czaplewski LG, North AK, Smith MC, Baumberg S, Stockley PG. (1992) Purification and initial characterization of AhrC: the regulator of arginine metabolism genes in Bacillus subtilis. Mol Microbiol. 6(2); 267-275.
JA. Garnett, S. Baumberg, PG. Stockley SEV Phillips (2007) A high-resolution structure of the DNA-binding domain of AhrC, the arginine repressor/activator protein from Bacillus subtilis Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007; 63: 914–917. Graphic prepared using PyMol.
Solving the expression & purification problems enabled solution of the structure of this interesting bi-functional gene regulatory protein.