Related Experiment Video
Updated: Aug 15, 2026
Long-Read Plasmid Sequencing and Assembly Using Nanopore Sequencing-Based Workflows
Published on: July 7, 2026
Determination of nucleotide sequence related to the plasmid replication region in Enterococcus faecalis and its
K Kirimura1, K Kamigaki, T Ebihara
1Department of Applied Chemistry, School of Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 169-8555, Japan.
Abstract:
The 5.1-kb plasmid pAMalpha1delta2, a derivative of the 9.6-kb plasmid pAMalpha1 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 pAMalpha1delta2 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 pAMalpha1delta2 and the replication regions of pAMalpha1delta2 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.
