Difference between revisions of "Part:BBa K902060:Experience"
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
how you used this part and how it worked out. | how you used this part and how it worked out. | ||
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| + | <p>In order to determine if the KatG-LAA protein was functional, catalase activity was tested using a turbitity comparison between <i>E. coli</i> expressing native catalase levels (carrying only the <i>pLacI</i> biobrick) and cultures of <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K902060"> <i>pLacI-katG-LAA</i></a>. Cultures were grown overnight, induced with 100 uM IPTG, and subsequently subcultured into various concentrations of hydrogen peroxide (0 mM, 1 mM, 5 mM, and 10 mM). These cultures were then grown overnight. It was found that the negative control exhibited no growth after 12h at 1 mM peroxide, however cultures with induced expression of catalase were turbid after 12 h of growth at this concentration (Fig. 3). This demonstrated the ability of the catalase to protect the cells from excessive peroxide concentrations.</p><p> | ||
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| + | </html>[[File:J04500-K137068 KatG assay sulfurucalgary.png|center|600px|thumb|Figure 3: Catalase Assay. Overnight cultures of pLacI and pLacI-KatGLAA were innoculated into 0 mM, 1 mM, 5 mM, and 10 mM peroxide. Cultures were grown overnight and turbidity was observed. It was found that at 1 mM of peroxide, cultures with just the lacI promotor perished, however when KatG-LAA was expressed, the cells survived.]]<html></p> | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 03:34, 3 October 2012
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Experience
In order to determine if the KatG-LAA protein was functional, catalase activity was tested using a turbitity comparison between E. coli expressing native catalase levels (carrying only the pLacI biobrick) and cultures of pLacI-katG-LAA. Cultures were grown overnight, induced with 100 uM IPTG, and subsequently subcultured into various concentrations of hydrogen peroxide (0 mM, 1 mM, 5 mM, and 10 mM). These cultures were then grown overnight. It was found that the negative control exhibited no growth after 12h at 1 mM peroxide, however cultures with induced expression of catalase were turbid after 12 h of growth at this concentration (Fig. 3). This demonstrated the ability of the catalase to protect the cells from excessive peroxide concentrations.
