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

[Reshma P. Shetty]

We do have a help forum on OWW.  :)

See http://openwetware.org/wiki/Questions_and_Answers

Joseph, I would post your question there so that it reaches a wider  
OWW audience.

-Reshma

PS Also see, http://openwetware.org/wiki/ 
Bacterial_artificial_chromosomes
Some of the BACs listed there have loxP sites.  But you have to be  
careful about which one you use to really get copy number 1-2.

On Sep 7, 2006, at 7:28 AM, tk at csail.mit.edu wrote:

> 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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>> your Internet provider.
>
>
>
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