Isolation and characterization of the rml gene homologs from Porphyromonas gingivalis

Yukie Shibata, Yoshihisa Yamashita, Y. Nakano, Toshihiko Koga

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We cloned four genes from the Porphyromonas gingivalis chromosome, the gene products of which catalyze the anabolism of dTDP-L-rhamnose from D-glucose-1-phosphate when they were expressed in Escherichia coli. The amino acid sequences deduced from these genes showed significant homology to proteins encoded by the rml genes involved in dTDP-L-rhamnose biosynthesis in other gram-negative bacteria. Reverse transcriptase polymerase chain reaction analysis revealed that these four genes are expressed as a single transcript in P. gingivalis. To clarify the role of the rml gene homologs in this organism, construction of mutants defective in the rml gene homologs was attempted by allelic exchange. Unexpectedly, any mutants defective in the rml gene homologs were unable to be isolated, and the allelic exchange was possible only if the wild-type rml gene homologs were present on the chromosome. These results suggest that the rml gene homologs might be essential for the viability of P. gingivalis under the culture conditions used in this study.

Original languageEnglish
Pages (from-to)339-347
Number of pages9
JournalOral Microbiology and Immunology
Volume14
Issue number6
DOIs
Publication statusPublished - Dec 1999

Fingerprint

Porphyromonas gingivalis
Genes
Chromosomes
Gram-Negative Bacteria
Reverse Transcriptase Polymerase Chain Reaction
Amino Acid Sequence
Escherichia coli
Glucose

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Dentistry(all)
  • Microbiology (medical)

Cite this

Isolation and characterization of the rml gene homologs from Porphyromonas gingivalis. / Shibata, Yukie; Yamashita, Yoshihisa; Nakano, Y.; Koga, Toshihiko.

In: Oral Microbiology and Immunology, Vol. 14, No. 6, 12.1999, p. 339-347.

Research output: Contribution to journalArticle

@article{f25977132fa14dd4846e5ba1a16d4997,
title = "Isolation and characterization of the rml gene homologs from Porphyromonas gingivalis",
abstract = "We cloned four genes from the Porphyromonas gingivalis chromosome, the gene products of which catalyze the anabolism of dTDP-L-rhamnose from D-glucose-1-phosphate when they were expressed in Escherichia coli. The amino acid sequences deduced from these genes showed significant homology to proteins encoded by the rml genes involved in dTDP-L-rhamnose biosynthesis in other gram-negative bacteria. Reverse transcriptase polymerase chain reaction analysis revealed that these four genes are expressed as a single transcript in P. gingivalis. To clarify the role of the rml gene homologs in this organism, construction of mutants defective in the rml gene homologs was attempted by allelic exchange. Unexpectedly, any mutants defective in the rml gene homologs were unable to be isolated, and the allelic exchange was possible only if the wild-type rml gene homologs were present on the chromosome. These results suggest that the rml gene homologs might be essential for the viability of P. gingivalis under the culture conditions used in this study.",
author = "Yukie Shibata and Yoshihisa Yamashita and Y. Nakano and Toshihiko Koga",
year = "1999",
month = "12",
doi = "10.1034/j.1399-302X.1999.140602.x",
language = "English",
volume = "14",
pages = "339--347",
journal = "Molecular Oral Microbiology",
issn = "2041-1006",
publisher = "American Journal of Nursing Company",
number = "6",

}

TY - JOUR

T1 - Isolation and characterization of the rml gene homologs from Porphyromonas gingivalis

AU - Shibata, Yukie

AU - Yamashita, Yoshihisa

AU - Nakano, Y.

AU - Koga, Toshihiko

PY - 1999/12

Y1 - 1999/12

N2 - We cloned four genes from the Porphyromonas gingivalis chromosome, the gene products of which catalyze the anabolism of dTDP-L-rhamnose from D-glucose-1-phosphate when they were expressed in Escherichia coli. The amino acid sequences deduced from these genes showed significant homology to proteins encoded by the rml genes involved in dTDP-L-rhamnose biosynthesis in other gram-negative bacteria. Reverse transcriptase polymerase chain reaction analysis revealed that these four genes are expressed as a single transcript in P. gingivalis. To clarify the role of the rml gene homologs in this organism, construction of mutants defective in the rml gene homologs was attempted by allelic exchange. Unexpectedly, any mutants defective in the rml gene homologs were unable to be isolated, and the allelic exchange was possible only if the wild-type rml gene homologs were present on the chromosome. These results suggest that the rml gene homologs might be essential for the viability of P. gingivalis under the culture conditions used in this study.

AB - We cloned four genes from the Porphyromonas gingivalis chromosome, the gene products of which catalyze the anabolism of dTDP-L-rhamnose from D-glucose-1-phosphate when they were expressed in Escherichia coli. The amino acid sequences deduced from these genes showed significant homology to proteins encoded by the rml genes involved in dTDP-L-rhamnose biosynthesis in other gram-negative bacteria. Reverse transcriptase polymerase chain reaction analysis revealed that these four genes are expressed as a single transcript in P. gingivalis. To clarify the role of the rml gene homologs in this organism, construction of mutants defective in the rml gene homologs was attempted by allelic exchange. Unexpectedly, any mutants defective in the rml gene homologs were unable to be isolated, and the allelic exchange was possible only if the wild-type rml gene homologs were present on the chromosome. These results suggest that the rml gene homologs might be essential for the viability of P. gingivalis under the culture conditions used in this study.

UR - http://www.scopus.com/inward/record.url?scp=0032743425&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032743425&partnerID=8YFLogxK

U2 - 10.1034/j.1399-302X.1999.140602.x

DO - 10.1034/j.1399-302X.1999.140602.x

M3 - Article

C2 - 10895688

AN - SCOPUS:0032743425

VL - 14

SP - 339

EP - 347

JO - Molecular Oral Microbiology

JF - Molecular Oral Microbiology

SN - 2041-1006

IS - 6

ER -