What is the function of thrombin in a plasmid? Does it help for screening, selection, or as a biomarker?
Dear Forbes
the thrombin site is not refer to the thrombin protein but to presence of a DNA sequence with codify for the recognition site for cleavage with thrombin.
Is quite common if you would like to fuse TAG (eg HIs, MBP, GST, Trx, GB1) to the N-term of your recombinant protein to improve its expression, solubity or purity, the addition of a protease (as thrombin, TEV, Fatt.Xa...) to allow you to remove those tags at the end of purifciation process.
On my blog: ProteoCool https://proteocool.blogspot.com/
at PAGE 2 you can find a video tutorial
ProteoCool n°22 (Overview on Tools for recombinant protein expression in E.coli )
where you can find a section dedicated to protease cleavage.
Ciao
Manuele
Dear Forbes
It is Protease cleavage site to separate and purify Bacterial Expressed protein of interest from fused tag like 6XHis, 4X or 10XMyc, GST and Trx.
Best Wishes
Thrombin is a protease. "Thrombin site" as labeled in the picture is the DNA sequence which contains the recognition site of thrombin. After purification of induced proteins carrying an affinity tag like 6-His/MBP/GST at their N-terminal end, thrombin is used to cleave the affinity tag.
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