Supplementary Figure 1: Analysis of MS2-tag insertion on growth at low temperature and incorporation into 70S ribosomes.

(a) Schematics of 16S rRNA indicating the positions of different MS2 tags in leader, trailer and mature sequence of 16S rRNA. The tags are explained in Fig. 1b in main text, in addition, tag 86L is 86nts into the Leader and between the RNase III and RNase E cleavage sites. (b) Growth of E. coli harboring different tagged and non-tagged, wild-type (WT) plasmids at non-permissive temperature (25°C). (c) Sucrose gradient sedimentation profiles of ribosomes and ribosomal subunits from wild-type strain carrying non-tagged (WT) or 105L tagged rDNA plasmid. Allele specific primer extension analysis of rRNA (for details, see Supplementary Fig. 2) from 30S and 70S ribosomal fractions collected from sucrose gradients reveals the plasmid borne 16S rRNA in each fraction and is plotted as a histogram (n = 3, standard deviation is shown as error bars).