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 4.5kb, 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.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Tagawa J,Inoue T,Naito M,Sato K,Kuwahara T,Nakayama M,Nakayama K,Yamashiro T,Ohara Ndoi
10.1016/j.mimet.2014.07.032subject
Has Abstractpub_date
2014-10-01 00:00:00pages
174-9eissn
0167-7012issn
1872-8359pii
S0167-7012(14)00228-0journal_volume
105pub_type
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