30S subunits from the reference strain MG 1655 were obtained by dissociation of 70S ribosomes. Briefly, 70S ribosomes purified according to [15] with minor modifications were dissociated into subunits under low magnesium concentration, fractionated in 15–30% (w/v) sucrose gradient, and the fractions containing 30S subunits were collected.

Immature ΔrbfA 30S particles from the ΔrbfA strain unable to associate with 50S subunits were purified by sucrose gradient centrifugation of the total ribosome fraction in regular salt conditions. The small ribosomal subunit fraction was considered to consist mainly of immature particles.

See Supplementary Materials for the detailed description of the procedures.

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