QUANTITATIVE-ANALYSIS OF RIBOSOME BINDING-SITES IN ESCHERICHIA-COLI

185 clones with randomized ribosome binding sites, from position - 11 to 0 preceding the coding region of beta-galactosidase, were selected and sequenced. The translational yield of each clone was determined; t hey varied by more than 3000-fold. Multiple linear regression analysis was used to determine the contribution to translation initiation acti vity of each base at each position. Features known to be important for translation initiation, such as the initiation codon, the Shine/Dalga rno sequence, the identity of the base at position - 3 and the occurre nce of alternative ATGs, are all found to be important quantitatively for activity. No other features are found to be of general significanc e, although the effects of secondary structure can be seen as outliers . A comparison to a large number of natural E. coli translation initia tion sites shows the information profile to be qualitatively similar a lthough differing quantitatively. This is probably due to the selectio n for good translation initiation sites in the natural set compared to the low average activity of the randomized set.