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Cheap procedure for gene deletion in E coli?


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#1 chemy333

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Posted 28 November 2010 - 12:20 PM

Hi,

I want to delete a particular gene from E coli to study if there is any effect on the phenotype etc. However, I don't want to introduce any antibiotic resistance, because the gene likely affects ribosome function so any antibiotics would interfere with followup experiments.

I've looked at the Red/ET recombination-based gene deletion kit by genebridges (http://www.genebridg...hp?prod_id=k006), which is really expensive. Do you know of a cheaper way to delete genes without getting additional antibiotic resistance?

Thanks

#2 perneseblue

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Posted 28 November 2010 - 08:09 PM

Hi,

I want to delete a particular gene from E coli to study if there is any effect on the phenotype etc. However, I don't want to introduce any antibiotic resistance, because the gene likely affects ribosome function so any antibiotics would interfere with followup experiments.

I've looked at the Red/ET recombination-based gene deletion kit by genebridges (http://www.genebridg...hp?prod_id=k006), which is really expensive. Do you know of a cheaper way to delete genes without getting additional antibiotic resistance?

Thanks


well you could recreate the system on your own, using Cre recombinase, FRT or phIC31. You could look around and request the plasmids (containing expressing enzymes) from other researchers.
May your PCR products be long, your protocols short and your boss on holiday

#3 pDNA

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Posted 29 November 2010 - 11:13 AM

you can go for the recombineering approach (pretty much the same like the pRED/ET system)

Recombineering: a homologous recombination-based method of genetic engineering

Regards,
p


Hi,

I want to delete a particular gene from E coli to study if there is any effect on the phenotype etc. However, I don't want to introduce any antibiotic resistance, because the gene likely affects ribosome function so any antibiotics would interfere with followup experiments.

I've looked at the Red/ET recombination-based gene deletion kit by genebridges (http://www.genebridg...hp?prod_id=k006), which is really expensive. Do you know of a cheaper way to delete genes without getting additional antibiotic resistance?

Thanks


well you could recreate the system on your own, using Cre recombinase, FRT or phIC31. You could look around and request the plasmids (containing expressing enzymes) from other researchers.






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