Optimized expression and purification of biophysical quantities of Lac repressor and Lac repressor regulatory domain.

ABSTRACT: The recombinant production of Lac repressor (LacI) in Escherichia coli is complicated by its ubiquitous use as a regulatory element in commercially-available expression vectors and host strains. While LacI-regulated expression systems are often used to produce recombinant LacI, the product can be heterogeneous and unsuitable for some studies. Alternative approaches include using unregulated vectors which typically suffer from low yield or vectors with promoters induced by metabolically active sugars which can dilute isotope labels necessary for certain biophysical studies. Here, an optimized expression system and isolation protocol for producing various constructs of LacI is introduced which eliminates these complications. The expression vector is an adaptation of the pASK backbone wherein

SUBMITTER: Stetz MA 

PROVIDER: S-EPMC4881282 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature