Hello, folks.
I am working in the characterization of a putative biosurfactant protein from Archaea. I screened out the gene from a metagenomics library, cloned it in expression vector with a His-tag at N terminal and over-expressed the recombinant protein with IPTG, at 37ºC. I've tried many protocols to visualize my over-expressed protein, purify it and I did not succeed. Oddly, when I tested the construction for other types of assays to assess surfactant activity I got impressive positive results, suggesting my surfactant protein is active and on fire. Despite this, I never achieved to visualize the superexpression in a SDS-PAGE dyed with Coomassie nor Silver Nitrate.
Can anyone help me with this issue? I would appreciate. Thank you very much.
OBS: This protein is probably secreted in the medium once is produced.
Thank you, Thamsanqa. I did not check inclusion bodies yet. Do you suggest/know a protocol to get proteins from inclusion bodies?
Besides doing what Thamsanqa suggests, run western blot using an anti-his antibody to determine the presence of your protein.
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