Date: Dec 12, 2009
Title: ATG:Newsletter 100120
-Assembly Cloning for Multi-Protein Expression
are waiting to be explored and developed:
- Gene Clusters of Protein Complexes of Eukaryotes and
- Artificial Biochemical Pathways
- Easy Modular Shuttle Vector Design
ACEMBL_IC for Insect Cells and ACEMBL-EC are available…
Please inquire for:
evoMAG - Expression Optimization by in silico Gene Evolution …
and our Cloning Service …
Figure 1. Most recent publications1,2 are confirming the observation the 5'-leader
sequence (indicated in green) is playing an important role in the transcriptional initiation by interacting with the
5'-end of the coding sequence in individual genes. This individuality of the coding sequence forming an individual
determinant of the different genes is the main reason for the necessity adapting genes by inclusion of the 5'-trailer sequence1.
In our example the constant Phage T7 trailer sequence T7_transl_en_RBS is available for example in
pET17b. This trailer is good
for a variety of genes but by far not for all. Please remember the initiation rate is the rate limiting step in translational processes
in case it is lower than the elongation rate. But in case the average elongation rate is higher than the translational initiation rate
other rules are becoming effective influencing parameters like polysomal package2 with impact on mRNA-stability, suppression of inclusion
bodies increase of the specific activity of the products and improvement of the general translationability of the genes optimized in this
1 Salis et al. (2009) Automated design of synthetic ribosome binding sites to control protein expression,Nature Biotechnology 27:946-950.
2 Brandt et al. (2009) The native 3D organization of bacterial polysomes. Cell 136:261-271.