Team:METU Turkey/Results Discussion/CooA Expression and Purification

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3-CooA Expression and Purification



1-Overexpression optimization
1. Expression vector comparison (pKK versus pTriEx)
When we compare the below results, the expression of pTriEx vector is higher than the expression of pTriEx vector.
10/09/18
Expression in BL21 with pTriEx
In this expression experiment, we obtained 2 different bands in BL21 pTriEx from BL21 negative control as indicated gel photo. CooA is found in two form (monomer and dimer).
10/10/05
Expression in JM109 and BL21 with pKK vector
In this expriment, for the first time we want to try CooA pKK vector in BL21. The vector was sent by Aono and he advised us to express it in JM109. But we have failed to express it. So we tried it in BL21. We have no result with JM109 and a little expression in BL21. Therefore; we gave up from expressing pKK vector in BL21. And JM109 grows slowly,also we never see any band in JM109 crude lysate in many unpublished data. Therefore; we lysated all bacteria with same OD600. But result wasn't changed. .