All experiments were performed with E. coli MG1655 (fnr+ rph-1) (Coli Genetic Stock Center #6300) and its isogenic derivatives. MG1655-ΔmoaA753::FRT-FRT was constructed by P1 transduction of the ΔmoaA753::FRT-kan-FRT allele from the Keio collection (44) into MG1655, followed by elimination of the kan marker using plasmid pCP20, which expresses flippase (FLP) recombinase (45). MG1655 derivatives carrying the ΔmoaA753::FRT-FRT, ΔdeoD780::FRT-FRT, Δapt754::FRT-FRT Δhpt743::FRT-FRT, and Δgpt756::FRT-kan-FRT alleles were likewise constructed by the addition of kan-containing gene deletions from the Keio collection followed by elimination of the kan marker using pCP20 on each iteration.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.