Ribosome Binding Site/about
The bacterial ribosome binds to particular sequences on an mRNA, primarily the Ribosome Binding Site (RBS) and the start codon (AUG). The RBS base-pairs with an RNA molecule that forms part of the bacterial ribosome (the 16s rRNA), while the start codon base-pairs with the initiator tRNA which is bound to the ribosome. In addition to the sequences of the RBS and the start codon being important, these two sequences need to be positioned approximately 6-7 nucleotides apart so they can both make contact with the appropriate parts of the ribosome complex. In the next section, we talk more about the specific interaction between the RBS and the ribosome.
The transfer curve of a BioBrick device is measured from input PoPs to output PoPs. For a protein-DNA inhibition system, the transfer curve depends on the rates and efficiencies of many stages in a chain such as transcriptional efficiency, mRNA lifetime, ribosome binding efficiency and several system-specific variables.
If BioBrick parts are to be interoperable, intentional engineering of this chain of events will be required to produce a desired transfer function.
An RBS is an RNA sequence upstream of the start codon that affects the rate at which a particular Open Reading Frame (ORF) is translated. Various aspects of RBS design affect the rate at which the ORF is translated. You can use this informtation either to better understand how RBSs work, or to design new RBSs. Before designing a new RBS, you should first see if you can use some of the existing high quality RBSs. This is because we think it is most useful to the community to develop a small number of very-well characterized collection of RBS rather than lots of rarely used RBSs.
synthesized rather than prepping a plasmid carrying the sequence from cells or attempting to amplify the RBS sequence via PCR. This tutorial briefly outlines the steps involved in making an RBS from de novo synthesized DNA.
|1. Design your RBS part||2. Order oligos||3. Double-stranding|
Detailed help on Ribosome Binding Sites
- Glossary of terms relating to RBS and translation initiation
- Issues related to RBSs and standard assembly.
- The consensus RBS for bacterial ribosomes and how it works