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andy_lord

Member Since 20 Feb 2013
Offline Last Active Feb 21 2013 05:01 PM
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Posts I've Made

In Topic: Very small peptide expression in E. coli

20 February 2013 - 02:23 AM

I have had some experience expressing a 14 aa peptide with both N-terminal His-tag and GST tag separately and I'd have to say it's much better expressing your peptide with a GST tag rather than a His-tag. My experience has taught me that GST being a highly soluble protein has better chances to "pull" your peptide to solubility than a normal 6X His-tag with a cleavage site.

But before you go for cloning and expressing a peptide this short I'd advice you to use a few bioinfo tools like DisEMBL or RONN to predict the disorder of your expressed peptide (the whole gene in case of the His-tag).

I'd agree with snowchild regarding detection and increased solubility and stability too. The shift in the gel for GST alone and GST tagged 16 aa peptide might be a little difficult to differentiate. Don't forget to use protease inhibitors. A peptide this small would be vulnerable to protease attacks.

Good luck!

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