Difference between revisions of "Part:BBa I712019:Experience"

theos

The Cambridge team confirmed that this part did emit light from E. coli when placed under an expression vector (:BBa_J13002) with the addition of D-luciferin. Despite the claim that the "part contains a Eukaryotic ribosome binding site and non-standard prefix and suffix" we were able to express it in E. coli after adding a prokaryotic RBS and promoter using standard BioBrick assembly.

Catramp

The Utah_State 2011 iGEM Team has confirmed that this part emits light when placed under control of a promoter and D-luciferin is added, and is capable of working with standard luciferase testing kits and luminometers.

Improvements: This part has been improved upon with the addition of Dual Luciferase Assay intermediates that make construction of promoter testing devices with this part much easier (Dual Luciferase Assay Component).

UT-Tokyo

UT-Tokyo 2011 Team has characterized that this part emits luminescence with a promoter and substrate (D-luciferin). For testing this device, we carried out a promoter assay using BBa_R0011 and BBa_J23119 as a control. We succesfully evaluated the relative expression levels of BBa_R0011 with various IPTG concentration, compared to that of BBa_J23119. For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page].

Improvements: This part has been utilized to devise a Firefly-Renilla Dual Luciferase Assay Kit. This kit makes an evaluation of promoters and other cis-elements much easyer and more quantitative. We also conducted a calibration of Photinus pyralis luciferase using a standard reagent, demonstrating that intracellular luciferase amount can be directly estimated from relative luminescence unit (RPU) using a linear regression. For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page].