Development of a novel plasmid vector pTIO-1 adapted for electrotransformation of Porphyromonas gingivalis

Junpei Tagawa, Tetsuyoshi Inoue, Mariko Naito, Keiko Sato, Tomomi Kuwahara, Masaaki Nakayama, Koji Nakayama, Takashi Yamashiro, Naoya Ohara

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We report here the construction of a plasmid vector designed for the efficient electrotransformation of the periodontal pathogen Porphyromonas gingivalis. The novel Escherichia coli-. Bacteroides/. P. gingivalis shuttle vector, designated pTIO-1, is based on the 11.0-kb E. coli-. Bacteroides conjugative shuttle vector, pVAL-1 (a pB8-51 derivative). To construct pTIO-1, the pB8-51 origin of replication and erythromycin resistance determinant of pVAL-1 were cloned into the E. coli cloning vector pBluescript II SK(-) and non-functional regions were deleted. pTIO-1 has an almost complete multiple cloning site from pBluescript II SK(-). The size of pTIO-1 is 4.5. kb, which is convenient for routine gene manipulation. pTIO-1 was introduced into P. gingivalis via electroporation, and erythromycin-resistant transformants carrying pTIO-1 were obtained. We characterized the transformation efficiency, copy number, host range, stability, and insert size capacity of pTIO-1. An efficient plasmid electrotransformation of P. gingivalis will facilitate functional analysis and expression of P. gingivalis genes, including the virulence factors of this bacterium.

Original languageEnglish
Pages (from-to)174-179
Number of pages6
JournalJournal of Microbiological Methods
Volume105
DOIs
Publication statusPublished - Oct 2014

Keywords

  • Electroporation
  • Periodontal disease
  • Plasmid
  • Porphyromonas
  • Transformation

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

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