Biotechnology Letters

, Volume 29, Issue 3, pp 431–437 | Cite as

A novel high-copy plasmid, pEC, compatible with commonly used Escherichia coli cloning and expression vectors

Original research paper

Abstract

A 66164-bp cryptic plasmid, pEIB1, was isolated from strain Vibrio anguillarum MVM425 and sequenced. A plasmid carrying a 1089-bp fragment, containing the minimal replication region of pEIB1, a kanamycin-resistance marker and an l-arabinose promoter, designated pEC, was maintained as a high copy plasmid in E. coli and stably inherited in the absence of antibiotic selection. Significantly, pEC was compatible with the widely used ColE1, pSC101 and p15A replicons making it a useful tool for a dual-plasmid expression system.

Keywords

Plasmid vector pEC Compatible origin Copy number Stability Dual plasmids expression Gene regulation 

References

  1. Atlung T, Brondsted L (1994) Role of the transcriptional activator AppY in regulation of the cyx appA operon of Escherichia coli by anaerobiosis, phosphate starvation, and grown phase. J Bacteriol 176:5414–5422PubMedGoogle Scholar
  2. Auger EA, Redding KE, Plumb I, Childs LC, Meng S-Y, Bennett GN (1989) Construction of lac fusions to the inducible arginine and lysine decarboxylase genes of Escherichia coli K12. Mol Microbiol 3:609–620PubMedCrossRefGoogle Scholar
  3. Chang ACY, Ncohen S (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the p15A cryptic miniplasmid. J Bacteriol 134:1141–1156PubMedGoogle Scholar
  4. del Solar G, Giraldo R, Ruiz-Echevarria MJ, Espinosa M, Diaz-Orejas R (1998) Replication and control of circular bacterial plasmids. Microbiol Mol Biol Rev 62:434–464PubMedGoogle Scholar
  5. Donoghue DJ, Sharp PA (1978) Replication of colicin E1 plasmid DNA in vivo requires no plasmid-encoded proteins. J Bacteriol 133:1287–1294PubMedGoogle Scholar
  6. Fu JF, Ying SW, Liu ST (1997) Cloning and characterization of the ori region of pSW1200 of Erwinia stewartii: similarity with plasmid P1. Plasmid 38:141–147PubMedCrossRefGoogle Scholar
  7. Fuller RS, Funnell BE, Kornber A (1984) The danA protein complex with the E. coli chromosomal replication origion (oriC) and other DNA site. Cell 38:889–900PubMedCrossRefGoogle Scholar
  8. Gale EF (1940) The production of amines by bacteria. 1. The decarboxylation of amino acids by strains of Bacterium coli. Biochem J 34:392–413PubMedGoogle Scholar
  9. Gallegos M-T, Michan C, Ramos JL (1993) The XylS/AraC family of regulators. Nucl Acid Res 21:807–810CrossRefGoogle Scholar
  10. Lee N (1980) Molecular aspects of ara regulation. In: Miller JHW, Reznikoff S (eds) The operon. Cold Spring Harbor Laboratory Press, New York, pp 389–410Google Scholar
  11. Lerner CG, Inouye M (1990) Low copy number plasmids for regulated low-level expression of cloned genes in Escherichia coli with blue/white insert screening capability. Nucl Acids Res 18:4631PubMedCrossRefGoogle Scholar
  12. Lin SY, Wu HZ, Ye J, Zhang HZ (2006) Construction of inducible promoter system in E.coli. J Shanghai Normal Univ (Nat Sci) 35:75–79Google Scholar
  13. Miller JH (1972) Experiments in molecular genetics. Cold spring Harbor Laboratory Press, New York, pp 352–355Google Scholar
  14. Nishihara K, Kanemori M, Kitagawa M, Yanagi H, Yura T (1998) Chaperone coexpression plasmids: differential and synergistic roles of DnaK–DnaJ–GrpE and GroEL–GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2 in Escherichia coli. Appl Environ Microbiol 64:1694–1699PubMedGoogle Scholar
  15. Pollock D, Bauer CE, Scolnik PA (1988) Transcription of the Rhodobacter capsulatus nifHDK operon is modulated by the nitrogensource. Construction of plasmid expression vectors based on the nifHDK promoter. Gene 65:269–275PubMedCrossRefGoogle Scholar
  16. Schleif RS (1992) DNA looping. Annu Rev Biochem 61:199–223PubMedCrossRefGoogle Scholar
  17. Shine J, Dalgarno L (1974) The 39-terminal sequence of E. coli 16S ribosomal RNA: complementary to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci 71:1342–1346PubMedCrossRefGoogle Scholar
  18. Stim-Herndon KP, Flores TM, Bennett GN (1996) Molecular characterization of adiY, a regulatory gene which affects expression of the biodegradative acid-induced arginine decarboxylase gene (adiA) of Escherichia coli. Microbiology 142:1311–1320PubMedCrossRefGoogle Scholar
  19. Stormo GD, Schneider TD, Gold TM (1982) Characterization of translational initiation sites in E. coli. Nucl Acids Res 10:2971–2996PubMedCrossRefGoogle Scholar
  20. Thomas CM, Stalker DM, Helinski DR (1981) Replication and incompatibility properties of segments of the origin region of replication of the broad host range plasmid RK2. Mol Gen Genet 181:1–7PubMedCrossRefGoogle Scholar
  21. Thomas CM, Cross MA, Hussain AAK, Smith CA (1984) Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2. EMBO J 3:57–63PubMedGoogle Scholar
  22. Tsutsui H, Fujiyama A, Murotsu T, Matsubara K (1983) Role of nine repeating sequences of the mini-F genome for expression of F-specific incompatibility phenotype and copy number control. J Bacteriol 155:337–344PubMedGoogle Scholar
  23. Veldscholte J, Ris-Stalpers C, Kuiper GG, Jenster G, Berrevoets C, Claassen E, van Rooij HC, Trapman J, Brinkmann AO, Mulder E (1990) A mutation in the ligand binding domain of the androgen receptor of human LNCaP cells affects steroid binding characteristics and response to anti-androgens. Biochem Biophys Res Commun 173:534–540PubMedCrossRefGoogle Scholar
  24. Vieira J, Messing J (1982) The pUC plasmids, an M13-derived system for insertion mutagenesis and sequencing with synthetic universal primer. Gene 19:259–268PubMedCrossRefGoogle Scholar
  25. Wang RF, Kushner SR (1991) Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli. Gene 100:195–199PubMedCrossRefGoogle Scholar
  26. Wu H, Ma Y, Zhang Y, Zhang H (2004) Complete sequence of virulence plasmid pEIB1 from the marine fish pathogen Vibrio anguillarum strain MVM425 and location of its replication region. J Appl Microbiol 97:1021–1028PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  1. 1.State Key Laboratory of Bioreactor Engineering (Mailbox 431)East China University of Science and TechnologyShanghaiChina

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