I am currently working with E. coli MG1655. I want to knock out the lacZ gene in the chromosome. Does anyone know a quick way to knock out this gene without using any phage? Thank you very much!
There are dozens of different ways to do it. However numerous lac deleted versions of MG1655 have been constructed and you can just get them directly from the stock center or from another lab. There are undoubtedly labs at your institution that have such a strain.
Hi Dr. Benedik, thank you so much for the answer. The reason I want to knock out lacZ is that this strain we have has some other genetic markers/knock out. Could you please introduce me one way that could be easily done? I have done konckout in BW25113 but not in MG1655, I am not sure with the red lambda recombination if I could use the same method. Thank you!
Yes you can use exactly the same method with MG1655 as you use for BW25113, just introduce the red recombination plasmids into that strain. Alternately if you know how to do P1 phage transductions you can also transduce a lacZ deletion using a linked marker.
Hi there...
I think transforming the MG1655 cells with pKD46 or pKD78 plasmid (codes for recombinase under pBAD promoter) should do your work.
- Badges
- Science topic