JM101 for Protein Expression?

[RayHL]

Hi everyone,

I'm planning some protein expressions and right now, I'm choosing my expression strains. I'm wondering about the ability of JM101 cells to express recombinant proteins. At my old lab, we used them for some expressions (and it did work fine), but after checking the genotype, they don't seem to lack proteases like ompT or lon, and they don't seem to have T7 oR T5 RNA polymerase.

So, the question is: why did they work at all? Or was it just a labeling error and we never really used JM101?

Thanks for your answers in advance!


Ray
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[David-Paul Minde]

not all proteins are proteolytically unstable (e.g. GFP is rather robust
once it is folded)
most proteins are easy to degrade, however, because there is usually some
flexible part which is easily accessible for proteolysis ...
D
2009/10/21 <RayHL@...>

> Hi everyone,
>
> I'm planning some protein expressions and right now, I'm choosing my
> expression strains. I'm wondering about the ability of JM101 cells to
> express recombinant proteins. At my old lab, we used them for some
> expressions (and it did work fine), but after checking the genotype, they
> don't seem to lack proteases like ompT or lon, and they don't seem to have
> T7 oR T5 RNA polymerase.
>
> So, the question is: why did they work at all? Or was it just a labeling
> error and we never really used JM101?
>
> Thanks for your answers in advance!
>
>
> Ray
> --
> Jetzt kostenlos herunterladen: Internet Explorer 8 und Mozilla Firefox 3.5
> -
> sicherer, schneller und einfacher! http://portal.gmx.net/de/go/chbrowser
>
> _______________________________________________
> Methods mailing list
> Methods@...
> http://www.bio.net/biomail/listinfo/methods
>

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[Jayakumar, R]

 
That shouldn't be a problem for expression in E.coli as long as it is recognized by native RNA polymerases.  Remember the plasmid antibiotic genes used for selection is recognized and expressed, so will other e.coli promoters used to express your gene.  For T7 to work, you will need an E.coli strain with the T7 phage genome as a lysogen expressing the viral T7 RNA polymerase, such as XL1blue-DE3.  
  best of luck
Jay

   
-----Original Message-----
From: methods-bounces@... [mailto:methods-bounces@...] On Behalf Of RayHL@...
Sent: Wednesday, October 21, 2009 8:36 AM
To: methods@...
Subject: JM101 for Protein Expression?

Hi everyone,

I'm planning some protein expressions and right now, I'm choosing my expression strains. I'm wondering about the ability of JM101 cells to express recombinant proteins. At my old lab, we used them for some expressions (and it did work fine), but after checking the genotype, they don't seem to lack proteases like ompT or lon, and they don't seem to have T7 oR T5 RNA polymerase.

So, the question is: why did they work at all? Or was it just a labeling error and we never really used JM101?

Thanks for your answers in advance!


Ray
--
Jetzt kostenlos herunterladen: Internet Explorer 8 und Mozilla Firefox 3.5 - sicherer, schneller und einfacher! http://portal.gmx.net/de/go/chbrowser

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