TY - JOUR
T1 - Determination of nucleotide sequence related to the plasmid replication region in Enterococcus faecalis and its application to a new shuttle vector
AU - Kirimura, Kohtaro
AU - Kamigaki, Kiyotake
AU - Ebihara, Tadashi
AU - Ishii, Yoshitaka
AU - Kanayama, Shinji
AU - Usami, Shoji
PY - 1999/1/1
Y1 - 1999/1/1
N2 - 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.
AB - 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.
KW - Electroporation
KW - Enterococcus faecalis
KW - Gelatinase
KW - Replication protein
KW - Shuttle vector
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U2 - 10.1016/S1389-1723(99)80115-7
DO - 10.1016/S1389-1723(99)80115-7
M3 - Article
C2 - 16232519
AN - SCOPUS:0033049506
SN - 1389-1723
VL - 87
SP - 566
EP - 571
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
IS - 5
ER -