How to exchange antibiotic resistance markers on plasmids for E
(by fulvio.celsi from gmail.com)
Tue Jun 16 10:36:00 EST 2009
why not simply move the gene instead of the resistance? easier....
Sector of Neurobiology,
International School for Advanced Studies, Scuola Internazionale di Studi
Superiori Avanzati (SISSA),
2009/6/16 David Liu <amanosz from yahoo.com>
> Has anyone exchanged or heard of someone exchanging the antibiotic
> resistance marker on a plasmid for E. coli? I need to co-transform E coli
> with two plasmids that have the same resistance marker, and the plasmids are
> not available with any other marker. I have plasmid sequence information
> for the two plasmids of interest and a 3rd plasmid with the new resistance
> marker that identifies the location of each marker. I've confirmed with a
> BLAST search that the annotated sequences are simply the coding sequences
> for each marker (start to stop codon). Is it simply a modular procedure
> where I remove one ORF and replace it with the other? Or are there other
> considerations I need to think about?
> The plasmid map doesn't annotate any promoter or terminator sequence for
> the resistance marker. But I assume if one is necessary, then I am only
> substituting the coding sequence and the remaining un-annotated architecture
> is still present.
> For what it's worth, I'd like to substitute a chloramphenicol resistance
> marker with a kanamycin marker.
> Thanks in advance for your help,
> Methods mailing list
> Methods from net.bio.net
More information about the Methods