IGEM:Caltech/2007/Project/cisTransResults: Difference between revisions
No edit summary |
|||
Line 22: | Line 22: | ||
The fluorescence levels of D1210, YFP (the YFP construct with no cis repressor), and YFP + aTc serve as a control (as in the case of D1210, to show the fluorescence levels of background cells), as a guideline for minimum activation needed (as titering results show that N and Q levels in the (no-cis) construct are sufficient to cause lysis), and as maximum protein production possible (100% level set by the YFP + aTc results). Thus, the YFP fluorescence levels provide a estimate of target trans activation levels. Arabinose is added to the trans samples to induce trans production, and aTc added to increase production of the cis transcript.<br> | The fluorescence levels of D1210, YFP (the YFP construct with no cis repressor), and YFP + aTc serve as a control (as in the case of D1210, to show the fluorescence levels of background cells), as a guideline for minimum activation needed (as titering results show that N and Q levels in the (no-cis) construct are sufficient to cause lysis), and as maximum protein production possible (100% level set by the YFP + aTc results). Thus, the YFP fluorescence levels provide a estimate of target trans activation levels. Arabinose is added to the trans samples to induce trans production, and aTc added to increase production of the cis transcript.<br> | ||
[[Image: | [[Image:20071020_cis3_2FINAL.jpg|right]] | ||
<br><br><br> | <br><br><br> | ||
<b>cis3 and accompanying trans</b> | <b>cis3 and accompanying trans</b> |
Latest revision as of 16:13, 26 October 2007
Quanta Data and AnalysesSo far, trans activation has been tested for cis3 and cis4 (with their trans combinations 1 and 2), as well as trans3 with cis8, making the total count:
The fluorescence levels of D1210, YFP (the YFP construct with no cis repressor), and YFP + aTc serve as a control (as in the case of D1210, to show the fluorescence levels of background cells), as a guideline for minimum activation needed (as titering results show that N and Q levels in the (no-cis) construct are sufficient to cause lysis), and as maximum protein production possible (100% level set by the YFP + aTc results). Thus, the YFP fluorescence levels provide a estimate of target trans activation levels. Arabinose is added to the trans samples to induce trans production, and aTc added to increase production of the cis transcript.
|
Summary
The most promising results are for cis3 and its trans combinations, or cis3trans1 and cis3trans2, which show significant trans activation. The next step involves transforming the trans1 and trans2 plasmids into the cis3-containing Q construct, and titering. Hopefully, trans activation levels are enough to give plaques for the cis3trans1 and cis3trains2, thus showing that Q levels are high enough for successful phage infection.