Immunogenic mycobacterial ribosomal and ribonucleic Acid preparations: chemical and physical characteristics.

Infection and Immunity
A S Youmans, G P Youmans

Abstract

Five to 20% linear sucrose gradients were used to obtain sedimentation patterns of mycobacterial ribosomes, ribosomal subfractions, and ribonucleic acid (RNA) preparations. Classical 70S ribosomes were obtained when 10(-1)m magnesium chloride was used. These, when dialyzed against 10(-4)m MgCl(2), yielded typical 50S, 30S, and smaller ribosomal subunits. The 30S subunits were the most immunogenic under these conditions. A ribosomal preparation containing subunits which varied from 2.5 to 40S was fractionated by collecting five fractions from a sucrose gradient; based upon the amount of nucleic acid present, the fraction containing the 40S particles was most immunogenic. Physical and chemical evidence suggested that mycobacterial RNA preparations extracted with 65% ethyl alcohol from the ribosomes and diluted in distilled water, were either double-stranded, or mostly double-helical, or had a highly organized secondary structure. This was based on the following observations. (i) Native RNA was resistant to trace amounts of ribonuclease. (ii) The approximate T(m) value in SSC buffer (0.15 m NaCl plus 0.015 m sodium citrate) was greater than 85 C and in 0.1 SSC buffer was 55 C; the RNA diluted in SSC buffer produced a hypochromic e...Continue Reading

Citations

Sep 1, 1978·CRC Critical Reviews in Microbiology·M Solotorovsky, M Lynn
Jan 1, 1977·Transactions of the Royal Society of Tropical Medicine and Hygiene·N GrymbergA O Lima
Dec 1, 1974·Bacteriological Reviews·F M Collins
Nov 1, 1972·Infection and Immunity·A S Youmans, G P Youmans
Apr 1, 1980·Infection and Immunity·M D CooperD V Bowser
Dec 1, 1982·Infection and Immunity·R L Gregory, I L Shechmeister

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