[OWW-SC] Vector with LoxP sites flanking one/two antibiotic markersfor e.coli

[tk@csail.mit.edu]

You might want to look at the Invitrogen "Gateway" vectors, which use Cre-Lox
recombination for moving inserts among the plasmids in their vector series.

This is probably not the right place to be asking this question.  I don't know
if we have a good forum site (uh, why not, folks?), but I frequent the one at
www.protocol-online.org, and asking this question again in their molecular
biology forum would likely yield better answers.  I'd also be happy to discuss
the matter in private email.  Just a quick note -- some of the common lab
strains such as TOP10 and DH10B have rpsL genotypes.  It's also a common and
easy mutation.


Quoting joseph borg <acidmucin at yahoo.co.uk>:

> Dear All,
>    
>   I am writing you to see whether there is anyone who is able to help me
> around.
>    
>   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.
>    
>   Thanks!
>    
>   Joseph
>    
>   Laboratory of Molecular Genetics
>   Department of Physiology and Biochemistry
>   Biomedical Science Building
>   University of Malta
>   Msida MSD06
>   MALTA
>    
>   tel: +356 2340 2774
>   fax: +356 2134 3535
>    
>    
>    
>    
>    
>    
> 
>  		
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