"
Team:Paris Liliane Bettencourt/Notebook/2011/08/12/
From 2011.igem.org
Danyel & Camille
tRNA
Due to uncertainties as to the eventual funcionality of the tRNA amber suppressor, we have decided to make several constructs:
1A) promotor Pveg + RBS (spoVG) + tRNA + Ter (B0015) 1B) Pveg + RBS + tRNA
P hyperspank + RBS + tRNA + Ter P hyperspank + RBS + tRNA
However, we have abandoned the constructs with the hyperspank promotor, because we received the wrong part from the registry.
Though ideally it would be best to not have an RBS preceding the gene, all promotor bricks already have an RBS attached to it. We have also verified that there is no ATG or TTG (the most used start codons in B.subtilis : "Translation in Bacillus subtilis: roles and trends of initiation and termination, insights from a genome analysis" Eduardo P. C. Rocha, Antoine Danchin and Alain Viari). There is a GTG codon (the least used start codon) at the 14 nucleotide of the gene sequence.
The part we received in place of the hyperspank promotor is a Pveg + RBS (different from spoVG). We decided to build two constructs with the same logic as the 1A) and 1B). They are temporarily named 4A) and 4B)
All parts were appropriately digested and are ready for ligation.
