JM101 for Protein Expression?
(by R.Jayakumar from roswellpark.org)
Wed Oct 21 13:06:17 EST 2009
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
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of RayHL from gmx.de
Sent: Wednesday, October 21, 2009 8:36 AM
To: methods from magpie.bio.indiana.edu
Subject: JM101 for Protein Expression?
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!
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