Team:METU Turkey/Results Discussion/CooA Expression and Purification
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<br>In expression comparison with IPTG concentration experiment, we realized that our CooA pTriEx vector is expressed without IPTG induction however when the cultures were induced with IPTG the expression incresed a little. Thus, we concluded that IPTG concentration doesn't much affect our expression yield. However, when 2mM IPTG was added to culture,expression of CooA was decreased. It has probably a toxic affect on our bacteria. | <br>In expression comparison with IPTG concentration experiment, we realized that our CooA pTriEx vector is expressed without IPTG induction however when the cultures were induced with IPTG the expression incresed a little. Thus, we concluded that IPTG concentration doesn't much affect our expression yield. However, when 2mM IPTG was added to culture,expression of CooA was decreased. It has probably a toxic affect on our bacteria. | ||
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+ | <br>D. Ferric Citrate effect | ||
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+ | <br>Time Dependent Expression of pTriEx vector with Ferric Citrate and without Ferric Citrate | ||
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+ | <br>10/10/10 | ||
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+ | <br>In the comparison of the cultures with Ferric Citrate and without Ferric Citrate, we prepared different cultures that all of them induced with IPTG and 4 of them had Ferric Citaret adn 4 of them did not have Ferric Citrate. According to the SDS-Page result, Ferric Citrate increses the expression of CooA. It is probably releated to heme characteristic of CooA. In this expreiment we also determine the best IPTG induction time. The cultures were induced for 3-5-7-9 hours. At the end of the experiment we concluded that the induction time also affects expression of CooA, however 5 hour induction is the best because both monomer and dimer form of CooA is highly expressed and the impurities are less when compared to 9 hours induction. In -FC7 well, we probably made pipeting error; therefore, we had greyed bands. | ||
Revision as of 18:15, 27 October 2010
3-CooA Expression and Purification
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