Questions and Answers
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Vector LoxP sites flanking two antibiotic marker genes for use in E.coli
I am in need of a low-copy vector (1 to 2 per cell) which carries two antibiotic marker genes. (preferably one sensitive such rpsL, and one resistant such at Tetracycline, Chloramphenicol, etc.) and are FLANKED between two LoxP sites so as once i express Cre recombinase i get recombination between the lox sites and removal of the antibiotic sites. I have rpsL gene at the lab, also i have tetracycline. So even if the plasmid doesn';t carry any antibiotic genes, but has LoxP it would be o.k and i may move in with my construct by subcloning between the sites but that would entail more work... :) Anyone in a position to help? I would also be very happy,whoever wishes,to collaborate and explain better my underlying project. email address: firstname.lastname@example.org
Is there an online or otherwise easy to use tool for finding RBS sites?
I've used TIGR's software to do RBS prediction. The program is called RBS finder. Also, if you have a specific sequence you want to test, you can use Stormo's work:
- Barrick D, Villanueba K, Childs J, Kalil R, Schneider TD, Lawrence CE, Gold L, and Stormo GD. Quantitative analysis of ribosome binding sites in E.coli. Nucleic Acids Res. 1994 Apr 11;22(7):1287-95.
Of couse, this is all for E. coli --Sri Kosuri 19:22, 23 March 2006 (EST)
Homologous recombination in E. coli
If anyone has experience with homologous recombination in E. coli please check out the Standard E. coli Strain for BioBricks project and annotate the page with your feedback and comments. Please include any protocols or materials information you might have on the appropriate pages and link to them from the standard strain project page as well. Any and all advice is appreciated.