Request for the ReDirect kit should be made to james.lipscombe@earlham.ac.uk. Request for cosmids in the original cosmid library should also be made to james.lipscombe@earlham.ac.uk. Cosmid names in the original cosmid library begin with "St". See here for more information.
ReDirect is a procedure developed in the JIC for knocking out genes in Streptomyces coelicolor. The procedure is generic and may be used on a variety of organisms.
If you are working on Streptomyces coelicolor then you can use the primer selection software specially developed for the ReDirect procedure.
23 June 2006: Now you can design primers which will allow you flip out the cassette by restriction digestion. Download BMW now for this feature.
A major bug which caused inaccurate embl file to be written for the disrupted cosmid has been fixed. If you are using a verion of BMW which was downloaded before November 2, 2004 then please down load the bugfixed version from here
25 May 2007: Juanpa has repackaged BMW for the Mac. Please read the readme file before downloading the image (dmg) file.
The protocol for the ReDirect procedure version 1.4 (in pdf format) is here and a powerpoint file showing new templates available for use in PCR targeted mutagenesis is here. The pdf of a manuscript entitled "Recombineering in Streptomyces coelicolor " is available here. Users of the ReDirect procedure might also find this paper useful.
A tool to (virtually) restrict cosmids is here.
In the S. coelicolor features display, the small red arrowheads indicate the positions of Tn5062 insertions. These insertions have been created in relevant cosmids that can be introduced into S. coelicolor strains by intergeneric conjugation from E. coli. The cosmids cannot replicate in S. coelicolor, but selection for the antibiotic resistance conferred by the transposon permits isolation of mutants arising by double crossover gene replacement (Bishop et al. 2004; Fernandez-Martinez et al. 2010).
These cosmids, each carrying a single unique Tn5062 insertion, as well as non-mutagenised cosmids spanning regions not covered by the original S. coelicolor cosmid library, can be obtained on request by emailing Meirwyn Evans (m.c.evans@swansea.ac.uk) and/or Paul Dyson (p.j.dyson@swansea.ac.uk).