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. 1996 Jun;178(12):3601–3607. doi: 10.1128/jb.178.12.3601-3607.1996

NBU1, a mobilizable site-specific integrated element from Bacteroides spp., can integrate nonspecifically in Escherichia coli.

N B Shoemaker 1, G R Wang 1, A A Salyers 1
PMCID: PMC178132  PMID: 8655560

Abstract

NBU1 is an integrated Bacteroides element that can he mobilized from Bacteroides donors to Bacteroides recipients. Previous studies have shown that a plasmid carrying the internal mobilization region of NBU1 could be transferred by conjugation from Bacteroides thetaiotaomicron to Escherichia coli. In this report, we show that NBU1 can integrate in E. coli. Whereas integration of NBU1 in B. thetaiotaomicron is site specific, integration of NBU1 in E. coli was relatively random, and the insertion frequency of NBU1 into the E. coli chromosome was 100 to 1,000 times lower than the frequency of integration in B. thetaiotaomicron. The frequency of NBU1 integration in E. coli could be increased about 10- to 70-fold, to a value close to that seen with B. thetaiotaomicron, if the primary integration site from B. thetaiotaomicron, BT1-1, was provided on a plasmid in the E. coli recipient or the NBU1 integrase gene, intN1, was provided on a high-copy-number plasmid to increase the amount of integrase available in the recipient. When the primary integration site was available in the recipient, NBU1 integrated site specifically in E. coli. Our results show that NBUs have a very broad host range and are capable of moving from Bacteroides spp. to distantly related species such as E. coli. Moreover, sequence analysis of NBU1 integration sites provided by integration events in E. coli has helped to identify some regions of the NBU1 attachment site that may play a role in the integration process.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bedzyk L. A., Shoemaker N. B., Young K. E., Salyers A. A. Insertion and excision of Bacteroides conjugative chromosomal elements. J Bacteriol. 1992 Jan;174(1):166–172. doi: 10.1128/jb.174.1.166-172.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  4. Bringel F., Van Alstine G. L., Scott J. R. Conjugative transposition of Tn916: the transposon int gene is required only in the donor. J Bacteriol. 1992 Jun;174(12):4036–4041. doi: 10.1128/jb.174.12.4036-4041.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kinder S. A., Badger J. L., Bryant G. O., Pepe J. C., Miller V. L. Cloning of the YenI restriction endonuclease and methyltransferase from Yersinia enterocolitica serotype O8 and construction of a transformable R-M+ mutant. Gene. 1993 Dec 22;136(1-2):271–275. doi: 10.1016/0378-1119(93)90478-l. [DOI] [PubMed] [Google Scholar]
  7. Kolter R., Inuzuka M., Helinski D. R. Trans-complementation-dependent replication of a low molecular weight origin fragment from plasmid R6K. Cell. 1978 Dec;15(4):1199–1208. doi: 10.1016/0092-8674(78)90046-6. [DOI] [PubMed] [Google Scholar]
  8. Komine Y., Adachi T., Inokuchi H., Ozeki H. Genomic organization and physical mapping of the transfer RNA genes in Escherichia coli K12. J Mol Biol. 1990 Apr 20;212(4):579–598. doi: 10.1016/0022-2836(90)90224-A. [DOI] [PubMed] [Google Scholar]
  9. Li L. Y., Shoemaker N. B., Salyers A. A. Characterization of the mobilization region of a Bacteroides insertion element (NBU1) that is excised and transferred by Bacteroides conjugative transposons. J Bacteriol. 1993 Oct;175(20):6588–6598. doi: 10.1128/jb.175.20.6588-6598.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Li L. Y., Shoemaker N. B., Wang G. R., Cole S. P., Hashimoto M. K., Wang J., Salyers A. A. The mobilization regions of two integrated Bacteroides elements, NBU1 and NBU2, have only a single mobilization protein and may be on a cassette. J Bacteriol. 1995 Jul;177(14):3940–3945. doi: 10.1128/jb.177.14.3940-3945.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Metcalf W. W., Jiang W., Wanner B. L. Use of the rep technique for allele replacement to construct new Escherichia coli hosts for maintenance of R6K gamma origin plasmids at different copy numbers. Gene. 1994 Jan 28;138(1-2):1–7. doi: 10.1016/0378-1119(94)90776-5. [DOI] [PubMed] [Google Scholar]
  12. Miller V. L., Mekalanos J. J. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Olsen G. J., Woese C. R., Overbeek R. The winds of (evolutionary) change: breathing new life into microbiology. J Bacteriol. 1994 Jan;176(1):1–6. doi: 10.1128/jb.176.1.1-6.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  15. Salyers A. A. Bacteroides of the human lower intestinal tract. Annu Rev Microbiol. 1984;38:293–313. doi: 10.1146/annurev.mi.38.100184.001453. [DOI] [PubMed] [Google Scholar]
  16. Shoemaker N. B., Getty C., Guthrie E. P., Salyers A. A. Regions in Bacteroides plasmids pBFTM10 and pB8-51 that allow Escherichia coli-Bacteroides shuttle vectors to be mobilized by IncP plasmids and by a conjugative Bacteroides tetracycline resistance element. J Bacteriol. 1986 Jun;166(3):959–965. doi: 10.1128/jb.166.3.959-965.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shoemaker N. B., Li L. Y., Salyers A. A. An unusual type of cointegrate formation between a Bacteroides plasmid and the excised circular form of an integrated element (NBU1). Plasmid. 1994 Nov;32(3):312–317. doi: 10.1006/plas.1994.1070. [DOI] [PubMed] [Google Scholar]
  18. Shoemaker N. B., Salyers A. A. A cryptic 65-kilobase-pair transposonlike element isolated from Bacteroides uniformis has homology with Bacteroides conjugal tetracycline resistance elements. J Bacteriol. 1990 Apr;172(4):1694–1702. doi: 10.1128/jb.172.4.1694-1702.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shoemaker N. B., Salyers A. A. Tetracycline-dependent appearance of plasmidlike forms in Bacteroides uniformis 0061 mediated by conjugal Bacteroides tetracycline resistance elements. J Bacteriol. 1988 Apr;170(4):1651–1657. doi: 10.1128/jb.170.4.1651-1657.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shoemaker N. B., Wang G. R., Salyers A. A. The Bacteroides mobilizable insertion element, NBU1, integrates into the 3' end of a Leu-tRNA gene and has an integrase that is a member of the lambda integrase family. J Bacteriol. 1996 Jun;178(12):3594–3600. doi: 10.1128/jb.178.12.3594-3600.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shoemaker N. B., Wang G. R., Stevens A. M., Salyers A. A. Excision, transfer, and integration of NBU1, a mobilizable site-selective insertion element. J Bacteriol. 1993 Oct;175(20):6578–6587. doi: 10.1128/jb.175.20.6578-6587.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith C. J., Parker A. C. Identification of a circular intermediate in the transfer and transposition of Tn4555, a mobilizable transposon from Bacteroides spp. J Bacteriol. 1993 May;175(9):2682–2691. doi: 10.1128/jb.175.9.2682-2691.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stevens A. M., Sanders J. M., Shoemaker N. B., Salyers A. A. Genes involved in production of plasmidlike forms by a Bacteroides conjugal chromosomal element share amino acid homology with two-component regulatory systems. J Bacteriol. 1992 May;174(9):2935–2942. doi: 10.1128/jb.174.9.2935-2942.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stevens A. M., Shoemaker N. B., Salyers A. A. The region of a Bacteroides conjugal chromosomal tetracycline resistance element which is responsible for production of plasmidlike forms from unlinked chromosomal DNA might also be involved in transfer of the element. J Bacteriol. 1990 Aug;172(8):4271–4279. doi: 10.1128/jb.172.8.4271-4279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stokes H. W., Hall R. M. A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: integrons. Mol Microbiol. 1989 Dec;3(12):1669–1683. doi: 10.1111/j.1365-2958.1989.tb00153.x. [DOI] [PubMed] [Google Scholar]
  26. Valentine P. J., Shoemaker N. B., Salyers A. A. Mobilization of Bacteroides plasmids by Bacteroides conjugal elements. J Bacteriol. 1988 Mar;170(3):1319–1324. doi: 10.1128/jb.170.3.1319-1324.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ward J. M., Grinsted J. Physical and genetic analysis of the Inc-W group plasmids R388, Sa, and R7K. Plasmid. 1982 May;7(3):239–250. doi: 10.1016/0147-619x(82)90005-1. [DOI] [PubMed] [Google Scholar]
  28. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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