Hello Everyone,
I am trying to develop a protein expression and purification system. Hence, can anyone tell me about "Intein Tags" or related papers, articles or suggestion?
Please Help
Thanks
Good places to start -
Paulus H. (2000) - Annu. Rev. Biochem. 2000. 69:447–96
Banki & Wood (2005) - http://microbialcellfactories.biomedcentral.com/articles/10.1186/1475-2859-4-32
Esipov et al (2008) - Protein Expression and Purification 61 (2008) 1–6
I have PDFs for all of these on my literature laptop so if you cannot obtain copies, please get in touch directly.
As other tags, Intein tag can be used at both C-terminus or N-terminus. Its dependent on what vector you choose. Intein tag can be used when you do not want any extra amino acids on your expressed protein. Cleaving from chitin column is simple; you can use 50mM DTT in column buffer. There are some ready made kits available to clone your GOI to the respective intein vectors.
Good Luck!
I've previously worked with intein-based purifications because of my PI's collaboration with David Wood (https://apel.osu.edu/). I suggest reviewing some of his publications for an introduction to the subject.
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