Difference between revisions of "Experience gained on new Registry parts"

Revision as of 00:55, 28 October 2008

Cell-to-cell signaling by XOR gate constructs - The ability of two different XOR gate constructs to send a signal via the Lux system was measured by coplating bacterial XOR clones with bacteria containing a Lux receiver construct.

Positive control using a Lux sender and a Lux receiver (K09100). Green fluorescence indicates LuxI expression in the Lux sender and receipt of the intercellular signal by the receiver.

XOR gate construct K091173 coplated with the K09100 Lux receiver. Green fluorescence indicates that at least one of the two hybrid promoters in the XOR construct is driving expression of the LuxI gene.

XOR gate construct K091182 coplated with the K09100 Lux receiver. Green fluorescence indicates that at least one of the two hybrid promoters in the XOR construct is driving expression of the LuxI gene.

Improved function by a modified Lac promoter - Mutations were introduced into the wild type Lac promoter that exists in the Registry. One mutant promoter, designated pLacIQ1, had improved ability to promote, as demostrated by the following experiment.

Constructs were assembled with eiter the wild type Lac promoter or mutant promoters driving GFP expression via part number E0240. Fluorescence measurements taken at 16 or 24 hours were corrected for optical density.

16 hours

24 hours

Sample Flourescence OD Flourescence/OD LB + Amp 1 0.047 21.27659574 E0240 3 1.201 2.497918401 S04020 4 1.239 3.228410008 S04021 14.5 1.216 11.92434211 K091131 71.3 1.207 59.07207954