Maximizing purification of ArnA?

06 June 2016 1 2K Report

I'm attempting to prep and purify PAO1-derived ArnA in a receptive E. coli strain. No one in our lab has attempted to purify this protein. I'm wondering if anyone has any experience with it--ie, what buffers we should use, what salt concentrations it likes, other factors that will affect how well it preps and purifies?

Dominique Liger

Hi there,

I have no experience with ArnA purification but I know this protein because it is one of the major contaminants after NiNTA step because of its natural affinity for this matrix. So if NiNTA is the option for you, I would advice to express your ArnA in an E coli strain engineered in order to avoid this contamination. The following papers should help:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4086167/

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3127686/

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