How to exchange antibiotic resistance markers on plasmids for E coli

Fulvio Celsi via methods%40net.bio.net (by fulvio.celsi from gmail.com)
Tue Jun 16 10:36:00 EST 2009


Dear David
why not simply move the gene instead of the resistance? easier....

Fulvio Celsi
BsC,PhD
Sector of Neurobiology,
International School for Advanced Studies, Scuola Internazionale di Studi
Superiori Avanzati (SISSA),
Trieste
Italy

Mobile: +393286489131


2009/6/16 David Liu <amanosz from yahoo.com>

>
> Hello,
>
> 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,
>
> David
>
>
>
>
>
>
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