2 replies to this topic

[thitima]

Hi,
I would like to known about the replicon type which come from pGC blue plasmid (pGC Blue Cloning kits; Lucigen® Corporation). I want to do the co-expression protocol with colE1 replicon. Can I use pGC vector?
If not, Which vector that I have to use with colE1? Please help me.

Thank you,

Thitima
:>.

[pDNA]

Dear Thitima,
  
      it's a ColE1 origin. If you want to use it for co-expression in E. coli you might experience problems, but you can try it if the two plasmids u are using have different resistance genes on it ...than apply both antibiotics in the medium (maybe this will work). It's reported in the literature that two ColE1 plasmids can't be stable in one cell ...but we never observed that when using selection pressure.

Other replication origins that you can use are p15A (10-12 copies) or pSC101 (~5 copies) ...they are 100% compatible with ColE1.

Regards,
                p
  

thitima, on Nov 27 2009, 07:42 AM, said:

Hi,
I would like to known about the replicon type which come from pGC blue plasmid (pGC Blue Cloning kits; Lucigen® Corporation). I want to do the co-expression protocol with colE1 replicon. Can I use pGC vector?
If not, Which vector that I have to use with colE1? Please help me.

Thank you,

Thitima
:>.

[thitima]

pDNA, Thank you very much.

:> Thitima