Fig. 5
From: Engineered ribosomes with tethered subunits for expanding biological function
Selected orthogonal pair sequences and function in Ribo-T v2. a Top evolved orthogonal mRNA and 16S with predicted pairing. Selection round is noted by round 1 or round 2 to the right of each pair. n denotes number of isolated members with that sequence from the selection. b–e Orthogonal pair notation: Original orthogonal Ribo-T system denoted by v1, and x.y where x is o16S number and y is o-mRNA letter (pORTx.y plasmid name format). b Orthogonal expression of super folder green fluorescent protein (sf-gfp) in BL21(DE3)Δupp.+ pair: both o-rRNA and o-mRNA expressed, − pair: just o-mRNA expressed without cognate o-rRNA. Percent orthogonality is shown below column labels. A higher percentage value is desired, indicating a lower background expression of o-mRNA as compared to the expression with the cognate orthogonal rRNA. Error bars = 1SD of n = 3 independent experiments. The protein’s structure and details are listed to the right of the graph. c Orthogonal expression of Cm acetyltransferase (cat) in BL21(DE3)Δupp. Error bars = 1 standard error in IC50 curve fitting. The protein’s structure and details are listed to the right of the graph. d Orthogonal expression of N-glycosyltransferase of A. pleuropneumoniae (ApNGT) in BL21(DE3). The protein’s structure and details are listed to the right of the graph. e Orthogonal expression of Beta-galactosidase (LacZ) in BL21(DE3). The protein’s structure and details are listed to the right of the graph. Source data for b, c can be found in the Source Data file
