Should I just use one RBS or should I try a few different RBS in parallel?
My protein isn't being expressed, could it be because of the RBS I picked?
Discuss RBS secondary structure issues and point to some examples of what can go wrong
Some RBS are short and some are long, does the ribosome bind to the whole RBS?
Explain the difference between an RBS "part" and an RBS
Where should the spacing be between the RBS and the start codon?
In bacteria, the ribosome binding site needs to be located approximately 6-7 nucleotides upstream of the start codon for a gene. Conveniently, the mixed site formed by a BioBrick assembly of an RBS and a coding sequence produces the correct spacing between the RBS and the start codon.
What does the strength of an RBS mean?
If the RBS just serves as a recognition site for the ribosome, why do we need more than one RBS sequence? Well, a couple of reasons. For now, suffice it to say that the rate ribosomes bind and initiate translation from an RBS is dependent on the sequence of the RBS. The higher the rate that ribosomes initiate translation from an RBS, the stronger we say the RBS is. Varying RBS sequence is a powerful way to control the level of gene expression in your biological system.
