Comparison of lysyl-transfer ribonucleic acid species from vegetative cells and spores of Bacillus subtilis by methylated albumin-kieselguhr and reversed-phase chromatography.
Lysyl-transfer ribonucleic acid (tRNA) species from a spore-forming strain of Bacillus subtilis (168 trp2(-)) and an early blocked asporogenous mutant (spoA 12) were compared on reversed-phase and methylated albumin-kieselguhr columns. Lysyl-tRNA species from spores and the asporogenous mutant in stationary phase both exhibited altered chromatographic profiles compared to that of log-phase cells. The major peak in spore lysyl-tRNA species eluted later than that characteristic of vegetative cells, whereas the major peak of the lysyl-tRNA species from the asporogenous mutant in stationary phase eluted earlier. Although the early eluting lysyl-tRNA species was observable on methylated albumin columns, the late eluting peak was not detectable by that column technique. By using a shallower gradient on an RPC-2 column, the resolution of all lysyl-tRNA species increased. Several subspecies were revealed. The chromatographic comparisons clearly show that both the spore-forming strain and the asporogenous mutant undergo relative increases in different lysyl-tRNA species when grown to late stationary phase. No new species seem to be involved but rather altered amounts of minor species existing in log-phase cells. The experiments also demonstrate the usefulness of reversed-phase columns for such comparisons.
D000418:Albumins; D001412:Bacillus subtilis / Q000032:analysis* / Q000254:growth & development / Q000378:metabolism; D002247:Carbon Isotopes; D002845:Chromatography; D005827:Genetics, Microbial; D008239:Lysine / Q000378:metabolism; D008722:Methods; D009154:Mutation; D012329:RNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D012343:RNA, Transfer / Q000032:analysis* / Q000302:isolation & purification; D012822:Silicon Dioxide; D013045:Species Specificity; D013053:Spectrophotometry; D013170:Spores / Q000032:analysis* / Q000254:growth & development / Q000378:metabolism; D014316:Tritium
D000418:Albumins; D002247:Carbon Isotopes; D012329:RNA, Bacterial; D014316:Tritium; D012822:Silicon Dioxide; D012343:RNA, Transfer; D008239:Lysine