Difference between revisions of "JCA Arkin RBSFamilyPart2"

Latest revision as of 00:55, 10 February 2008

Characterization of the Ribosome Binding Site Library

To date, the RBS library has not been quantitatively characterized. This is largely a reflection of the method used to construct the library. Initiatially, all members of the library had the RBS variant upstream of an RFP gene. I measured red fluorescence for 384 individual library members, and the ranked activity of them is shown in the chart above. All screened clones showing activity detectable above background were combined into 7 activity pools. Each pool underwent a procedure designed to remove the RFP gene and put back the SpeI site in each part. 96 individual processed clones were then sequenced, and the family of RBS parts reflects all the unique clones.

So, at this stage we really can't say much about their relative activity. All I can tell you is that they should all initiate translation at a rate above background, and they are very roughly in decreasing order of activity. RBS's with low numbers are likely to be stronger RBS's than those with higher numbers.

Hopefully this will change in the near future, and if that happens the info. will be posted here.

...and of course YOU could do some characterization and post the info here.

The raw data for the library after sequenceing is Media:030807-Bca1050 raw rbs data.xls. The Tecan data of the clones before repooling / FokI fragmentation is Media:JCA_010607-TecanRBS.xls.

The original nomenclature was Bca1050 and then Bca9110.