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During this study His-tagged CcpA protein purified under native conditions to obtain a biologically active protein was used for molecular analysis of CcpA-dependent regulation. Using electrophoretic mobility shift assays it was demonstrated that CcpA of L. lactis can bind DNA in the absence of the HPr-Ser-P corepressor and exhibits DNA-binding affinity for nucleotide sequences lacking cre sites. However,...
In this work we present cloning and overexpression of lactococcal CcpA protein in Escherichia coli Xllblue strain as a fusion with 6xHis tag. A high yield of the CcpA protein was obtained when the cells were cultured in liquid medium LB with 100 ug/ml ampicillin at 37°C and subsequently for 4 h after induction by IPTG. The procedure let us obtain 5 mg of homogenous CcpA protein. Glutaraldehyde crosslinking...
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