Development of a novel plasmid vector pTIO-1 adapted for electrotransformation of Porphyromonas gingivalis.
J Microbiol Methods
; 105: 174-9, 2014 Oct.
Article
em En
| MEDLINE
| ID: mdl-25102110
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 4.5kb, 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.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Transformação Bacteriana
/
Técnicas de Transferência de Genes
/
Eletroporação
/
Porphyromonas gingivalis
/
Vetores Genéticos
/
Genética Microbiana
/
Biologia Molecular
Limite:
Humans
Idioma:
En
Revista:
J Microbiol Methods
Ano de publicação:
2014
Tipo de documento:
Article
País de afiliação:
Japão