+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Complete sequence of virulence plasmid pEIB1 from the marine fish pathogen Vibrio anguillarum strain MVM425 and location of its replication region



Complete sequence of virulence plasmid pEIB1 from the marine fish pathogen Vibrio anguillarum strain MVM425 and location of its replication region



Journal of Applied Microbiology 97(5): 1021-1028



The aim of this study was to determine the whole DNA sequence of pEIB1, one pJM1-like virulence plasmid from Vibrio anguillarum MVM425 and locate the replication region. DNA sequence of virulence plasmid pEIB1 from V. anguillarum MVM425 was determined using the methods of restriction endonuclease digestion, subcloning, and primer walking. The whole nucleotide sequence of pEIB1 comprises 66,164 bp, encoding 44 open reading frames (>400 bp) containing the genes of DNA replication, biosynthesis and regulation of the siderophore anguibactin and transport of ferric-anguibactin complexes. With no demonstrated replication origin, the Sau3AI partial digested plasmid DNA fragments of pEIB1 were ligated into the BamHI-fragment containing the kanamycin-resistance gene (Kmr). For there is no effective transformation in V. anguillarum, the ligated DNA was first introduced into E. coli JM83, and the transfomants were selected for resistance to kanamycin. It was demonstrated with southern blotting and DNA sequencing that plasmid pEIB7 containing the Sau3AI DNA fragment of pEIB1 (from 12516 to 13957) has the ability to replicate in E. coli JM83 and V. anguillarum MVM425sh. The segregational stability of plasmid pEIB7 kept in 100 and 4% in E. coli JM83 and V. anguillarum MVM425sh respectively when the cells were cultured in 200th generation. In following experiments, we also found that plasmid pEIB7 replicated at a middle-copy number of 10-40 in JM83, while at a high-copy number of 100-300 in MVM425sh. Moreover, pEIB7 can survive in V. alginolyticus, another fish pathogenic. With the whole DNA sequence of pEIB1 determining, it was found that pEIB1 showed microheterogeneity in its restriction endonuclease patterns with pJM1 though their DNA sequences had slight difference. According to the complete DNA sequence of pEIB1, its replication region was located from 12516 to 13957. And this replication region is compatible to pUC18 (pMB1), pKA3 (pSC101) and p15A: caiE (p15A). The worldwide vibriosis marine pathogen V. anguillarum strains contain common virulence, pJM1-like plasmids, independent on the geographical source. The pEIB1 was the second common virulence plasmid, which sequence was determined. Its sequence is highly homologous to pJM1 as they both encode biosynthesis and regulation of the siderophore anguibactin and transport of ferric-anguibactin complexes. Some interesting features as in pJM1 were also identified, such as transposon-like structures. So it can be deferred that the whole DNA sequences of virulent plasmid pEIB1 will be great helpful to future revealing these V. anguillarum virulence-related genes derived during evolution from transposition events or horizontal transfer of genes potentially originating in other organisms. Another result, replication region of pEIB1 locating is the first report about replication of pJM1-like plasmid. This work will be useful for researching pJM1-like plasmid replication mechanism in V. anguillarum.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 004084367

Download citation: RISBibTeXText

PMID: 15479418

DOI: 10.1111/j.1365-2672.2004.02387.x


Related references

Complete sequence of virulence plasmid pJM1 from the marine fish pathogen Vibrio anguillarum strain 775. Journal of Bacteriology 185(19): 5822-5830, 2003

Complete genome sequence of the marine fish pathogen Vibrio anguillarum harboring the pJM1 virulence plasmid and genomic comparison with other virulent strains of V. anguillarum and V. ordalii. Infection and Immunity 79(7): 2889-2900, 2011

Two replication regions in the pJM1 virulence plasmid of the marine pathogen Vibrio anguillarum. Plasmid 67(2): 95-101, 2012

Virulence plasmid pJM1 prevents the conjugal entry of plasmid DNA into the marine fish pathogen Vibrio anguillarum 775. Journal of General Microbiology 138(12): 2485-2490, 1992

DNA sequencing of a plasmid with virulence from marine fish pathogen Vibrio anguillarum. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica et Biophysica Sinica 35(10): 956-959, 2003

The genetics of plasmid mediated virulence in the marine fish pathogen vibrio anguillarum. Crosa, J H (Ed ) Bacterial And Viral Diseases Of Fish: Molecular Studies X+86p A Washington Sea Grant Publication: Seattle, Washington Illus Paper : p21-30, 1983

Curing of a plasmid is correlated with an attenuation of virulence in the marine fish pathogen Vibrio anguillarum. Infection and Immunity 27(3): 897-902, 1980

A plasmid associated with virulence in the marine fish pathogen Vibrio anguillarum specifies an iron-sequestering system. Nature 284(5756): 566-568, 1980

A plasmid associated with virulence in the marine fish pathogen Vibrio anguillarum specifies an iron-sequestering system. Nature, UK 284(5756): 566-568, 1980

Evidence for plasmid contribution to the virulence of the fish pathogen vibrio anguillarum. Abstracts Of The Annual Meeting Of The American Society For Microbiology: 0, 1977

Evidence for plasmid contribution to the virulence of fish pathogen Vibrio anguillarum. Infection and Immunity 18(2): 509-513, 1977

Complete sequence of plasmid pMP1 from the marine environmental Vibrio vulnificus and location of its replication origin. Marine Biotechnology 11(4): 456-462, 2008

Genetic Determinants of Virulence in the Marine Fish Pathogen Vibrio anguillarum. Fish Pathology 46: 1-10, 2011

Complete genome sequence of the marine fish pathogen Vibrio anguillarum and genome-wide transposon mutagenesis analysis of genes essential for in vivo infection. Microbiological Research 216: 97, 2018

Polypeptides p40 pom2 and pangr are required for iron uptake and for virulence of the marine fish pathogen of vibrio anguillarum 775. Journal of Bacteriology 173(3): 1347-1352, 1991