Molecular Characterization of Plasmid from Bifidobacterium longum

  • Park, Myeong-Soo (Research Center, BIFIDO Co., Ltd.) ;
  • Moon, Hye-Won (Department of Food Science and Technology, Seoul National University) ;
  • Ji, Geun-Eog (Research Center, BIFIDO Co., Ltd., Department of Food and Nutrition, Seoul National University)
  • Published : 2003.06.01

Abstract

The complete nucleotide sequence of a plasmid, pMG1, isolated from Bifidobacterium longum MG1 has been determined. This plasmid, composed of 3,862 base pairs with 65.1% of G+C content. harbors two major open reading frames (ORF) encoding putative proteins of 29 kDa (ORF I) and 71 kDa (ORF II). ORF I showed relatively high amino acid sequence homology with replication proteins of other plasmids from Gr Im-positive and -negative bacteria. Upstream of ORF I, four sets of tandem repeat sequences resembling the iteron structure of related plasmids were found. S1 endonuclease treatment and Southern blot analysis revealed that pMG1 accumulates single-stranded DNA (ssDNA) intermediate, which indicate i the rolling circle replication (RCR) mechanism of this plasmid. Homology search indicated that ORF II encodes plasmid mobilization protein, and the presence of highly conserved oriT sequence in the upstream of this gene supported this assumption. RT-PCR showed that only ORF I is expressed in vivo. Based on these results, pMG 1 was exploited to construct a shuttle vector, pBES2. It was successfully transformed into Bifidobacterium and maintained stably.

Keywords

References

  1. Microbiology v.142 A convenient and reproducible method to genetically transform bacteria of the genus Bifidobacterium Argnani,A.;R.J.Leer;N.vanLuijk;P.H.Pouwels https://doi.org/10.1099/13500872-142-1-109
  2. FEMS Microbiol. Lett. v.110 Analysis of the genome of five Bifidobacterium breve strains: Plasmid content, Pulsed-field gel electrophoresis genome size estimation and run loci number Bourget,N.;J.M.Simonet;B.Decaris https://doi.org/10.1111/j.1574-6968.1993.tb06288.x
  3. Res. Microbiol. v.152 Effects of probiotic administration upon the composition and enzymatic activity of human fecal microbiota in patients with irritable bowel syndrome or functional diarrhea Brigidi,P.;B.Vitali;E.Swennen;G.Bazzocchi;D.Matteuzzi https://doi.org/10.1016/S0923-2508(01)01254-2
  4. J. Bacteriol. v.178 Identification and characterization of the origin of conjugarive transfer (oriT) and a gene (nes) encoding a single-stranded endonuclease on the Staphylococcal plasmid pGO1 Climo,M.W.;V.K.Sharma;G.L.Archer
  5. J. Bacteriol. v.174 The oriT region of the Agrobacterium tumefaciens Ti plasmid pTiC58 shares DNA sequence identity with the transfer origins of RSF1010 and RK2/RP4 and with T-region borders Cook,D.M.;S.K.Farrand
  6. Appl. Microbiol. Biotechnol. v.53 Nucleotide sequence and thermostability of pND324, a3.6-kb plasmid from Lactococcus lactis Duar,K.;C.Q.Liu;Y.J.Liu;J.Ren;N.W.Dunn https://doi.org/10.1007/s002530051611
  7. Gene v.113 Replication and copy number control of the broad-hostrange plasmid RSF1010 Frey,J.;M.M.Bagdasarian;M.Bagdasarian https://doi.org/10.1016/0378-1119(92)90675-F
  8. Microbiol. Rev. v.53 The family of highly interrelated single stranded deoxyribonuleic acid plasmids Gruss,A.;S.D.Ehrlich
  9. Bifidobact. Microfl. v.5 Effects of fructooligosaccharides on intestinal flora and human health Hidaka,H.;T.Takizawa;T.Tokunaga;Y.Toshiro
  10. Methods Mol. Biol. v.25 CLUSTAL V:Multiple alignments of DNA and protein sequences Higgins,D.G.
  11. Am. J. Clin. Nutr. v.73 Probiotics in human disease Isolauri,E.
  12. Antonie Van Leeuwenhoek v.82 Discovering lactic acid bacteria by genomics Klaenhammer,T.;E.Altermann;F.Arigoni;A.Bolotin;F.Breidt;J.Broadbent;R.Cano;S.Chaillou;J.Deutscher;M.Gasson;M.van de Guchte;J.Guzzo;A.Hartke;T.Hawkins;P.Hols R.Hutkins;M.Kleerebezem;J.Kok;O.Kuipers;M.Lubbers;E.Maguin;L.McKay;D.Mills;A.Nauta;R.Overbeek;H.Pel;D.Pridmore;M.Saier;D.van Sinderen;A.Sorokin;J.Steele;D.O'Sullivan;W.de Vos;B.Weimer;M.Zagorec;R.Siezen https://doi.org/10.1023/A:1020638309912
  13. J. Microbiol. Biotechnol. v.11 Characterization of plasmids from Bifidobacterium sp. Lee,J.H.;M.S.Park;K.H.Lee;G.E.Ji
  14. FEMS Immunol. Med. Microbiol. v.10 Modulation of a specific humoral immune response and changes in intestinal flora mediated through fermented milk intake Link-Amster, H.;F.Rochat;K.Y.Saudan;O.Mignot;J.M.Aeschlimann https://doi.org/10.1111/j.1574-695X.1994.tb00011.x
  15. Lett. Appl. Microbiol. v.11 Characterization and molecular cloning of Bifidobacterium longum cryptic plasmid pMB1 Matteuzzi,D.;P.Brigidi;M.Rossi;D.Digioia https://doi.org/10.1111/j.1472-765X.1990.tb00165.x
  16. Plasmid v.32 Transformation of Bifidobacterium longum with pRM2, a constructed Escherichia coli-B. longum shuttle vector Missich,R.;B.Sgobati;D.J.LeBlanc https://doi.org/10.1006/plas.1994.1056
  17. Biosci. Biotechnol. Biochem. v.66 Cloned cytosine deaminase gene expression of Bifidobacterium longum and application to enzyme/pro-drug therapy of hypoxic solid tumors Nakamura,T.;T.Sasaki;M.Fujimori;K.Yazawa;Y.Kano;J.Amano;S.Taniguchi https://doi.org/10.1271/bbb.66.2362
  18. J. Microbiol. Biotechnol. v.11 Molecular cloning and characterization of the β-galactosidase gene from Bifidobacterium adolescentis Int57 Park,M.S.;H.J.Yoon;S.L.Rhim;G.E.Ji
  19. J. Microbiol. Biotechnol. v.10 Characterization of plasmid pKJ36 from Bifidobacterium longum and construction of an E. coli-Bifidobacterium shuttle vector Park,M.S.;D.W.Shin;K.H.Lee;G.E.Ji
  20. Microbiology v.145 Sequence analysis of plasmid pKJ50 from Bifidobacterium longum Park,M.S.;D.W.Shin;K.H.Lee;G.E.Ji https://doi.org/10.1099/13500872-145-3-585
  21. Lett. Appl. Microbiol. v.25 Isolation and characterization of two plasmids from Bifidobacterium longum Park,M.S.;K.H.Lee;G.E.Ji https://doi.org/10.1046/j.1472-765X.1997.00059.x
  22. Res. Microbiol. v.147 Characterization of the plasmid pMB1 from Bifidobacterium longum and its use for shuttle vector construction Rossi,M.;P.Bridigi;A.Gonzalez Vara y Rodriguez;D. Matteuzzi https://doi.org/10.1016/0923-2508(96)80213-0
  23. Lancet v.344 Feeding of Bifidobacterium bifidum and Streptococcus thermophilus to infants in hospital for prevention of diarrhoea and shedding of rotavirus Saavedra,J.M.;N.A.Bauman;I.Oung;J.A.Perman;R.H.Yolken https://doi.org/10.1016/S0140-6736(94)91708-6
  24. Molecular Cloning: A Laboratory Manual(2nd ed.) Sambrook,J.;E.F.Fritsch;T.Maniatis
  25. Mol. Gen. Genet. v.226 DnaA protein/DNA interaction. Modulation of the recognition sequence Schaefer,C.;W.Messer https://doi.org/10.1007/BF00273584
  26. Proc. Natl. Acad. Sci. USA v.99 The genome sequence of Bifidobacterium longum reflects its acaptation to the human gastrointestinal tract Schell,M.A.;M.Karmirantzou;B.Snel;D.Vilanova;B.Berger;G.Pessi;M.C.Zwahlen;F.Desiere;P.Bork;M.Delley;R.D.Pridmore;F.Arigoni https://doi.org/10.1073/pnas.212527599
  27. Microbiologica v.9 Related structures in the plasmid profiles of Bifidobacterium longum Sgorbati,B.;V.Scadovi;D.J.LeBlanc
  28. Nucleic Acid Res. v.20 Conserved sequence motifs in the initiator proteins for rolling circle DNA replication encoded by diverse replicons from eubacteria, eucaryotes and archaebacteria Tatyana,V.;V.K.Eugene https://doi.org/10.1093/nar/20.13.3279
  29. J. Bacteriol. v.177 Streptococcal plasmid pIP501 has a functional oriT site Wang,A.;L.Macrina