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. 2015 Aug 26;127(43):12982–12985. doi: 10.1002/ange.201506311

Figure 3.

Figure 3

rRNA derived from pRSF‐O‐ribo(h44H101) is assembled into functional ribosomes in vivo. A) Northern blot using a probe specific to the O‐ASD sequence detects RNAs from total RNA extracts that bear O‐ASDs. Wild‐type (WT) E.  coli, which does not possess orthogonal ribosomes, generates no band. O‐ribo possesses an O‐ASD on the 16S rRNA, and generates a band at 1500 nt (the length of 16S rRNA). pRSF‐O‐ribo(h44H101) generates an O‐ASD‐containing band near 4500 nt (nucleotides). B) Increasing isopropyl β‐d‐thiogalactopyranoside (IPTG) concentrations up to 0.1 mm increases the expression of orthogonal ribosomes (blue and black traces). Stable O‐ribo(h44H101) rRNA is generated to about 25 % the levels of O‐ribo (red trace). C) Growth curves of E.  coli bearing an O‐cat reporter, with or without O‐ribo(h44H101), at 37 °C in liquid LB media supplemented with IPTG and chloramphenicol (Cm).