Supplementary Figure 2: Outline for the purification of tagged SSU assembly intermediates from wild-type E. coli. | Nature Structural & Molecular Biology

Supplementary Figure 2: Outline for the purification of tagged SSU assembly intermediates from wild-type E. coli.

From: Multiple in vivo pathways for Escherichia coli small ribosomal subunit assembly occur on one pre-rRNA

Supplementary Figure 2

MS2CP = MS2 coat protein fused with Maltose Binding Protein (MBP). Plasmid borne 16S rRNA carry the spectinomycin resistance mutation, C1192U, which is absent in genomic operons. Allele specific primer extension of 16S rRNA purified with tags at position 105L, 11L and 20T using Cy5 labeled primer starting reverse transcription at position 1193 in the presence of ddGTP and differential stops are detected on 20% polyacrylamide gel. 16S represents mature 16S rRNA from genomic operons. Quantification shows more than 95% rRNA is plasmid borne. 1192C is chromosomally encoded while 1192U is plasmid encoded 16S rRNA.

Back to article page