Team:Groningen/project AND gate
From 2011.igem.org
AND gate
In order to integrate the memory units, we designed the AND gate system. To modulate gene expression at the translation level, we harness the use of engineered RNA-based/riboregulator system. After transcription, the 5’ UTR of crRNA-LasR transcript will form a stem-loop structure that blocks the RBS and prevents translation. To activate the expression of a self-repressed gene, the taRNA which is complementary to crRNA will unfold the stem-loop structure thus exposing the obstructed RBS and permit translation. (Isaacs, 2006)
Parts
The design of the AND gate circuit is based on two constructs, noncoding taRNA-DT cloned downstream of a hybB promoter (PhybB) and a cI promoter (PcI) cloned upstream crRNA-RBS-LasR-TAG-DT. In attempt to characterize the functional assembled parts, we cotransformed plasmid harboring AND gate system with reporter plasmid downstream LasR promoter.