Bacterial strains and transformation procedures
This protocol is extracted from research article:
Escherichia coli as a platform for the study of phosphoinositide biology
Sci Adv, Mar 27, 2019; DOI: 10.1126/sciadv.aat4872

For all cloning and plasmid maintenance procedures, we used the DH5α E. coli strain. While the system that we designed should work on any E. coli strain, since it uses only the endogenous protein production machinery, we performed all the experiments using E. coli strain BL21 DE3, which is commonly used for protein overexpression owing to the presence of T7 polymerase (53). We chose this widely used strain to show that the production of phosphoinositides could be easily controlled in it, since it is most familiar to other researchers and thus the most likely strain to be used with our system in the future. Bacteria were made chemically competent in our laboratory using the Mix & Go E. coli Transformation Kit from Zymo Research, which follows a standard chemical competence protocol.

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