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Criteria

View our judging criteria for iGEM 2011 here.

Wildtype

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.99566, 0.95671, 0.880328, 0.734557, 0.662629, 0.607491, 0.530352 0.012905, 0.015917, 0.020404, 0.026943, 0.013849, 0.0068952, 0.0060456 4

Mutant 1

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.6492, 0.20526, 0.17329, 0.16899, 0.16682, 0.16631, 0.16913 0.017682, 0.0032754, 0.0010138, 0.0011266, 0.00097033, 0.0012072, 0.0011179

Mutant 2

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.53303, 0.16064, 0.14171, 0.12149, 0.12547, 0.11146, 0.1061 0.02651, 0.0029121, 0.0027198, 0.0033112, 0.0034421, 0.0059837, 0.0027095

Mutant 4

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.23, 0.038825, 0.035663, 0.036119, 0.035423, 0.034289, 0.032858 0.02048, 0.00047265, 0.00028731, 0.00037973, 0.00031652, 0.00026622, 0.00070058

Mutant 5

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.92311, 0.23346, 0.1833, 0.17092, 0.16534, 0.16473, 0.15538 0.0061434, 0.0029621, 0.0016015, 0.0037633, 0.0029015, 0.002096, 0.0024785

Mutant 6

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.97182, 0.98592, 1.01000, 1.02300, 1.03610, 1.04490, 1.01750 0.018621, 0.01966, 0.018634, 0.018826, 0.018993, 0.020762, 0.019842

Mutant 7

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.48325, 0.13919, 0.12353, 0.10617, 0.10943, 0.099213, 0.08565 0.012073, 0.0025601, 0.0030506, 0.0027667, 0.002179, 0.0022998, 0.0015159

Mutant 8

SEQUENCE GOES HERE! 0, .0014, .0028, .0042, .0056, .0070, .0140 0.41657, 0.12325, 0.10744, 0.10365, 0.10006, 0.096006, 0.09219 0.0080566, 0.0028055, 0.0027614, 0.0038199, 0.0038188, 0.0024541, 0.0022523

The above widget displays the relative GFP fluorescence data that we have collected for the eight lambda cI mutant promoters and the wild type R0051 promoter. The plots are normalized to the wild type such that the wild type's fluorescence at 0% arabinose and 0 mM IPTG is 1. The data for these plots were gathered using the constructs outlined on the lambda cI project page, which were designed so that increasing the arabinose concentration would proportionally increase the repressor concentration. This widget allows you to toggle the display of whichever mutants you would like to view and easily observe the changes that these mutations cause.

Analysis

It is evident from the above plots that the variants in this lambda cI mutant library vary quite a bit in maximal promoter activity and in repressor sensitivity, especially when compared to wild type. In this data set, we see a very good range in activity level when there is minimal repressor present, but when the concentration of repressor is increased, the promoters quickly become fully repressed. This could mean that we have increased the binding affinity of these sequences to the lambda cI repressor, but we would need to examine sequences before we can know for sure. Additionally, with Mutant #6, it appears as though we have eliminated the ability of the repressor to bind all together. Once again, sequences for this part would be necessary to confirm this suspicious. Nonetheless, having these various levels of basal promoter activity is very useful and has many potential applications to synthetic biology.

Team:UC Davis/Data LambdacI

From 2011.igem.org