Determination of nucleotide sequence related to the plasmid replication region in Enterococcus faecalis and its application to a new shuttle vector

Kohtaro Kirimura, Kiyotake Kamigaki, Tadashi Ebihara, Yoshitaka Ishii, Shinji Kanayama, Shoji Usami

Research output: Contribution to journalArticle

Abstract

The 5.1-kb plasmid pAMα1Δ2, a derivative of the 9.6-kb plasmid pAMα1 which is harbored by Enterococcus faecalis ATCC 14508, has a region necessary for replication in E. faecalis. The nucleotide sequence related to the replication region in pAMα1Δ2 was determined and found to contain an open reading frame of 720-bp encoding a replication protein. The sequence showed 54.5 and 48.5% homology to those encoding the RepAs of plasmids pLA103 from Lactobacillus acidophilus and pFA3 from Neisseria gonorrhoeae, respectively. A recombinant 5.8-kb plasmid, pEFX6, which can be used as a shuttle vector between Escherichia coli and some strains of E. faecalis, was constructed by combining the tetracycline resistance gene of pAMα1 and the replication regions of pAMα1Δ2 and pUC18 for E. faecalis and E. coli, respectively. This shuttle vector was successfully used to clone and express the gelatinase gene from E. faecalis subsp. zymogenes IFO 3989 in E. faecalis C57, a strain showing no gelatinase activity.

Original languageEnglish
Pages (from-to)566-571
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume87
Issue number5
DOIs
Publication statusPublished - 1999 Jan 1

Keywords

  • Electroporation
  • Enterococcus faecalis
  • Gelatinase
  • Replication protein
  • Shuttle vector

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Determination of nucleotide sequence related to the plasmid replication region in Enterococcus faecalis and its application to a new shuttle vector'. Together they form a unique fingerprint.

  • Cite this