Which E.coli host strains can be used with the IMPACT vectors?
The fusion constructs should initially be established in a non-expression strain. Vector DNA supplied with the IMPACT system is prepared from a restriction-deficient E. coli strain (r-m-). When introduced into a strain with wild type EcoK (hsd+) DNA will be restricted. Thus plasmid DNA or ligated DNA should be introduced into a restriction-deficient E. coli strain (r-m- or r-m+). The IMPACT kit includes the non-competent NEB T7 expression strain ER2566. A competent version of ER2566, T7 Express Competent E. coli ( NEB #C2566H), can be purchased separately from New England Biolabs, Inc. or is available as a Special Offer with the IMPACT Kit ( NEB#E0543S). Several commercially available strains have also been tested, all of which allowed for expression and purification with the IMPACT system. Note: the culture should be inoculated with a freshly grown colony. For optimal expression of each fusion protein different strains should be tested. For E.coli strains to be used, go to expression
