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Contents 1 Construction 1.1 Plasmids backbones 1.2 Constructs for Synechocystis 2 Characterization in Synechocystis 2.1 Transformation 2.2 Verification of constructs 2.3 Bioluminescence Assays 2.3.1 Bioluminometer 2.3.2 High-sensitive camera 3 Experimental Highlights

Construction

Plasmids backbones

We built two integrative plasmid backbones for Synechocystis PCC. 6803.

Constructs for Synechocystis

Using our plasmid backbones we proceeded to build our constructs for Synechocystis:

Luciferase (LuxAB):

We tried many times (>6) to construct the substrate production (LuxCDEG) plasmid, however we were unsuccessful. We are currently designing another strategy to build the construct...

Characterization in Synechocystis

Transformation

We used our plasmid backbones to transform Synechocystis PCC. 6803.

Transformation with pSB1C3_IntK (BBa_K743006):

Transformation with pSB1C3_IntS (BBa_K743010):

Verification of constructs

We proceeded to verificate the integration of the constructs in Synechocystis.

(New image with illustration of verification from outer RS1 and RS2 - October 1X something)

Bioluminescence Assays

Bioluminometer

-graph-

High-sensitive camera

During the Latin America Jamboree, we had a chat with a couple of judges and a student, David Olarte, on how to induce Synechocystis with n-decanal. David sent us some work he had done on luminescence assays in cyanobacteria. Following his methods finally we were able to see light emition.

Experimental Highlights