What is this video? It's a narrated slide show by Jason Kelly introducing and detailing proposed technical standard for measuring the activity of engineered transcription promoters and ribosome binding sites in bacteria.
Wait wait wait! Whats a transcription promoter?!
Wiki Entry: In order for transcription to take place, the enzyme that synthesizes RNA, known as RNA polymerase, must attach to the DNA near a gene. Promoters contain specific DNA sequences and response elements which provide a binding site for RNA polymerase and for proteins called transcription factors that recruit RNA polymerase.
- In bacteria, the promoter is recognized by RNA polymerase and an associated sigma factor, which in turn are brought to the promoter DNA by an activator protein binding to its own DNA sequence nearby.
- In eukaryotes, the process is more complicated, and at least seven different factors are necessary for the binding of an RNA polymerase II to the promoter.
Promoters represent critical elements that can work in concert with other regulatory regions (enhancers, silencers, boundary elements/insulators) to direct the level of transcription of a given gene.
OK. What are ribosome binding sites?A ribosomal binding site (RBS) is a sequence on mRNA that is bound by the ribosome when initiating protein translation.It can be either the 5' cap of a messenger RNA in eukaryotes, a region 6-7 nucleotides upstream of the start codon AUG in prokaryotes (called the Shine-Dalgarno sequence), or an internal ribosome entry site (IRES) in viruses. The sequence is complementary to the 3' end of the rRNA. The ribosome searches for this site and binding to it (through base-pairing) begins the translation process, recruiting initiation factors.
This presentation was developed for the March 1 2008 BioBricks Foundation (BBF) Technical Standards Workshop held at UCSF.For more information see:
BBF Synth Bio Notes
Notes from the BBF meeting (BBF = Biobricks Foundation)
Registry/Measurement kit
BBF Synth Bio Protocols
/Promoter characterization experiment (FACS)
Make sure to visit: http://www.biobricks.org/
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