Isolation and amino acid sequence of the 30S ribosomal protein S19 fromMycobacterium bovisBCG

Naoya Ohara, Makoto Kimura, Yoshinori Higashi, Takeshi Yamada
1993 FEBS Letters  
The 30s ribosomal proteins from Mycobacteriurw bovis BCG were separated by reverse phase-high performance liquid chromatography (RP-HPLC). The isolated protems were analyzed by SDS-PAGE, blotted on PVDF-membranes and subjected to sequence analyses using a gas-phase sequencer to correlate them to those of the well studred Escher&a coli and Bucillus steurothernlophilus ribosomes. Moreover. the internal amino acid sequence of one ribosomal protem, MboS19, whtch IS homologous to E. colr rtbosomal
more » ... otein Sl9 (EcoS.19) and B. strurotlzermophrlus ribosomal protein S19 (BstSl9). was further analyzed by sequencing Its internal peptides and two segments from the N-and C-termini of the protein were selected to deduce the sequence of two ohgonucleotrde primers whtch were used m a polymerase chain reaction. Usmg the amplified DNA fragment thus obtained as a hybridization probe. the gene encoding protein S19 was identified and cloned. Sequence analysis of the DNA fragment, together with peptide sequence analysis could determine the complete ammo acid sequence of MboS19. This sequence proved to be 64% and 71% identical to those of the corresponding S19 proteins from the eubacteria E &I, and B. stearothern~ophrkts, respectively: 33% of the restdues of MboS19 were identical to those m the archaebacteral ribosomal protein HmaS19 Mycobacterww bows BCG; Ribosomal protein; Ammo actd sequence Published by Elsevier Science Publishers B V
doi:10.1016/0014-5793(93)80287-5 pmid:8405418 fatcat:4mxcrulbnfawtoovin6bfdctfi