Plant Molecular Biology

, Volume 23, Issue 4, pp 905–909 | Cite as

A conjugative plasmid vector for promoter analysis in several cyanobacteria of the genera Synechococcus and Synechocystis

  • Pierre Marraccini
  • Stéphane Bulteau
  • Corinne Cassier-Chauvat
  • Pierre Mermet-Bouvier
  • Franck Chauvat
Short Communication


A promoter-probe vector, pSB2A, based on the plasmid RSF1010 and the promoterless chloramphenicol acetyl transferase (cat) reporter gene, has been constructed. pSB2A appeared to be most efficiently transferred by conjugation to the widely used cyanobacteria Synechocystis strains PCC6803 (S.6803) and PCC6714 (S.6714) and Synechococcus strains PCC7942 (S.7942) and PCC6301 (S.6301), where it replicates stably even though it contains no cyanobacterial DNA. Using pSB2A we found that (1) a light-regulated promoter from S.6803 remains controlled by light intensity in S.7942 while it is silent in Escherichia coli, and (2) the E. coli tac promoter behaves as a strong and light-independent promoter in the four cyanobacterial hosts tested.

Key words

conjugation cyanobacteria light regulation promoter probe RSF1010 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Berkhout, B, Kastelein, RA, vanDuin, J: Translational interference at overlapping reading frames in prokaryotic messenger RNA. Gene 37: 171–189 (1985).CrossRefPubMedGoogle Scholar
  2. 2.
    Brosius, J: Plasmid vectors for the selection of promoters. Gene 27: 151–160 (1984).CrossRefPubMedGoogle Scholar
  3. 3.
    Brosius, J, Holly, A: Regulation of ribosomal RNA promoters with a synthetic lac operator. Proc Natl Acad Sci USA 81: 6929–6933 (1984).PubMedGoogle Scholar
  4. 4.
    Chang, ACY, Cohen, SN: Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the p15A cryptic plasmid. J Bact 134: 1141–1156 (1978).PubMedGoogle Scholar
  5. 5.
    Chauvat, F, deVries, L, van derEnde, A, vanArkel, GA: A host-vector system for gene cloning in the cyanobacterium Synechocystis PCC 6803. Mol Gen Genet 204: 185–191 (1986).CrossRefGoogle Scholar
  6. 6.
    DeBoer, HA, Comstock, LJ, Vasser, M: The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci USA 80: 21–25 (1983).PubMedGoogle Scholar
  7. 7.
    Elhai, J, Wolk, CP: Conjugal transfer of DNA to cyanobacteria. Meth Enzymol 167: 747–754 (1988).PubMedGoogle Scholar
  8. 8.
    Ferino, F, Chauvat, F: A promoter-probe vector-host system for the cyanobacterium Synechocystis PCC 6803. Gene 84: 257–266 (1989).CrossRefPubMedGoogle Scholar
  9. 9.
    Frey, J, Bagdasarian, M: The molecular biology of IncQ plasmids. In: Thomas, CM (eds) Promiscuous Plasmids of Gram-Negative Bacteria, pp. 79–94. Academic Press, London (1989).Google Scholar
  10. 10.
    Kuhlemeier, CJ, Thomas, AAM, van derEnde, A, vanLeen, RW, Borrias, WE, van denHondel, CAMJJ, vanArkel, GA: A host-vector system for gene cloning in the cyanobacterium Anacystis nidulans R2. Plasmid 10: 156–163 (1983).PubMedGoogle Scholar
  11. 11.
    Kreps, S, Ferino, F, Mosrin, C, Gerits, J, Mergeay, M, Thuriaux, P: Conjugative transfer and autonomous replication of a promiscuous IncQ plasmid in the cyanobacterium Synechocystis PCC6803. Mol Gen Genet 221: 129–133 (1990).CrossRefGoogle Scholar
  12. 12.
    Prentki, P, Krisch, HM: In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29: 303–313 (1984).CrossRefPubMedGoogle Scholar
  13. 13.
    Rippka, R, Deruelles, J, Waterbury, JB, Herdman, M, Stanier, RY: Generic assignments, strain histories and properties of pure culture of cyanobacteria. J Gen Microbiol 111: 1–61 (1979).Google Scholar
  14. 14.
    Scanlan, DJ, Bloye, SA, Mann, NH, Hodgson, DA, Carr, NG: Construction of lacZ promoter probe vectors for use in Synechococcus: application to the identification of CO2-regulated promoters. Gene 90: 43–49 (1990).CrossRefPubMedGoogle Scholar
  15. 15.
    Scholz, P, Haring, V, Wittman-Liebold, B, Ashman, K, Bagdasarian, M, Scherzinger, E: Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010. Gene 75: 271–288 (1989).CrossRefPubMedGoogle Scholar
  16. 16.
    Shaw, WV: Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. In: Hash, JE (eds) Methods in Enzymology, vol. 43, pp. 737–755. Academic Press, New York (1975).Google Scholar
  17. 17.
    Stueber, D, Bujard, H: Transcription from efficient promoters can interfere with plasmid replication and diminish expression of plasmid genes. EMBO J 1: 1399–1404 (1982).PubMedGoogle Scholar
  18. 18.
    Tandeau, N, Houmard, J: Adaptation of cyanobacteria to environmental stimuli: new steps towards molecular mechanism. FEMS Microbiol Rev 104: 119–190 (1993).CrossRefGoogle Scholar
  19. 19.
    Thiel, T, Poo, H: Transformation of a filamentous cyanobacterium by electroporation. J Bact 171: 5743–5746 (1989).PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Pierre Marraccini
  • Stéphane Bulteau
  • Corinne Cassier-Chauvat
    • 1
  • Pierre Mermet-Bouvier
  • Franck Chauvat
  1. 1.CNRS, URA D1290, Départment de Biologie Cellulaire et Moléculaire, Commissariat à l'Energie Atomiqu eCentre d'Etudes de SaclayGif-sur-Yvette CedexFrance

Personalised recommendations