Isolation and amino acid sequence of the 30S ribosomal protein S19 from Mycobacterium bovis BCG

Naoya Ohara, Makoto Kimura, Yoshinori Higashi, Takeshi Yamada

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The 30S ribosomal proteins from Mycobacterium bovis BCG were separated by reverse phase-high performance liquid chromatography (RP-HPLC). The isolated proteins 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 studied Escherichia coli and Bacillus stearothermophilus ribosomes. Moreover, the internal amino acid sequence of one ribosomal protein, MboS19, which is homologous to E. coli ribosomal protein S19 (EcoS19) and B. stearothermophilus ribosomal protein S19 (BstS19), 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 oligonucleotide primers which were used in a polymerase chain reaction. Using 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 amino acid sequence of MboS19. This sequence proved to be 64% and 71% identical to those of the corresponding S19 proteins from the eubacteria E. coli, and B. stearothermophilus, respectively; 33% of the residues of MboS19 were identical to those in the archaebacteral ribosomal protein HmaS19.

Original languageEnglish
Pages (from-to)9-14
Number of pages6
JournalFEBS Letters
Volume331
Issue number1-2
DOIs
Publication statusPublished - Sep 27 1993

Fingerprint

Geobacillus stearothermophilus
Mycobacterium bovis
Amino Acid Sequence
Ribosomal Proteins
Escherichia coli Proteins
Escherichia coli
Amino Acids
Proteins
DNA Primers
Protein Sequence Analysis
Protein C
DNA Sequence Analysis
Ribosomes
Peptides
Gene encoding
Sequence Analysis
Polyacrylamide Gel Electrophoresis
Polymerase chain reaction
DNA
High performance liquid chromatography

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Isolation and amino acid sequence of the 30S ribosomal protein S19 from Mycobacterium bovis BCG. / Ohara, Naoya; Kimura, Makoto; Higashi, Yoshinori; Yamada, Takeshi.

In: FEBS Letters, Vol. 331, No. 1-2, 27.09.1993, p. 9-14.

Research output: Contribution to journalArticle

Ohara, Naoya ; Kimura, Makoto ; Higashi, Yoshinori ; Yamada, Takeshi. / Isolation and amino acid sequence of the 30S ribosomal protein S19 from Mycobacterium bovis BCG. In: FEBS Letters. 1993 ; Vol. 331, No. 1-2. pp. 9-14.
@article{372ce17386684716b4d8df534976bcbc,
title = "Isolation and amino acid sequence of the 30S ribosomal protein S19 from Mycobacterium bovis BCG",
abstract = "The 30S ribosomal proteins from Mycobacterium bovis BCG were separated by reverse phase-high performance liquid chromatography (RP-HPLC). The isolated proteins 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 studied Escherichia coli and Bacillus stearothermophilus ribosomes. Moreover, the internal amino acid sequence of one ribosomal protein, MboS19, which is homologous to E. coli ribosomal protein S19 (EcoS19) and B. stearothermophilus ribosomal protein S19 (BstS19), 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 oligonucleotide primers which were used in a polymerase chain reaction. Using 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 amino acid sequence of MboS19. This sequence proved to be 64{\%} and 71{\%} identical to those of the corresponding S19 proteins from the eubacteria E. coli, and B. stearothermophilus, respectively; 33{\%} of the residues of MboS19 were identical to those in the archaebacteral ribosomal protein HmaS19.",
author = "Naoya Ohara and Makoto Kimura and Yoshinori Higashi and Takeshi Yamada",
year = "1993",
month = "9",
day = "27",
doi = "10.1016/0014-5793(93)80287-5",
language = "English",
volume = "331",
pages = "9--14",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Isolation and amino acid sequence of the 30S ribosomal protein S19 from Mycobacterium bovis BCG

AU - Ohara, Naoya

AU - Kimura, Makoto

AU - Higashi, Yoshinori

AU - Yamada, Takeshi

PY - 1993/9/27

Y1 - 1993/9/27

N2 - The 30S ribosomal proteins from Mycobacterium bovis BCG were separated by reverse phase-high performance liquid chromatography (RP-HPLC). The isolated proteins 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 studied Escherichia coli and Bacillus stearothermophilus ribosomes. Moreover, the internal amino acid sequence of one ribosomal protein, MboS19, which is homologous to E. coli ribosomal protein S19 (EcoS19) and B. stearothermophilus ribosomal protein S19 (BstS19), 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 oligonucleotide primers which were used in a polymerase chain reaction. Using 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 amino acid sequence of MboS19. This sequence proved to be 64% and 71% identical to those of the corresponding S19 proteins from the eubacteria E. coli, and B. stearothermophilus, respectively; 33% of the residues of MboS19 were identical to those in the archaebacteral ribosomal protein HmaS19.

AB - The 30S ribosomal proteins from Mycobacterium bovis BCG were separated by reverse phase-high performance liquid chromatography (RP-HPLC). The isolated proteins 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 studied Escherichia coli and Bacillus stearothermophilus ribosomes. Moreover, the internal amino acid sequence of one ribosomal protein, MboS19, which is homologous to E. coli ribosomal protein S19 (EcoS19) and B. stearothermophilus ribosomal protein S19 (BstS19), 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 oligonucleotide primers which were used in a polymerase chain reaction. Using 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 amino acid sequence of MboS19. This sequence proved to be 64% and 71% identical to those of the corresponding S19 proteins from the eubacteria E. coli, and B. stearothermophilus, respectively; 33% of the residues of MboS19 were identical to those in the archaebacteral ribosomal protein HmaS19.

UR - http://www.scopus.com/inward/record.url?scp=0027423321&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027423321&partnerID=8YFLogxK

U2 - 10.1016/0014-5793(93)80287-5

DO - 10.1016/0014-5793(93)80287-5

M3 - Article

C2 - 8405418

AN - SCOPUS:0027423321

VL - 331

SP - 9

EP - 14

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-2

ER -