Advertisement

Plant Cell Reports

, Volume 25, Issue 6, pp 582–591 | Cite as

Swapped green algal promoters: aphVIII-based gene constructs with Chlamydomonas flanking sequences work as dominant selectable markers in Volvox and vice versa

  • A. Hallmann
  • S. Wodniok
Genetic Transformation and Hybridization

Abstract

Production of transgenic organisms is a well-established, versatile course of action in molecular biology. Genetic engineering often requires heterologous, dominant antibiotic resistance genes that have been used as selectable markers in many species. However, as heterologous 5′ and 3′ flanking sequences often result in very low expression rates, endogenous flanking sequences, especially promoters, are mostly required and are easily obtained in model organisms, but it is much more complicated and time-consuming to get appropriate sequences from less common organisms. In this paper, we show that aminoglycoside 3′-phosphotransferase gene (aphVIII) based constructs with 3′ and 5′ untranslated flanking sequences (including promoters) from the multicellular green alga Volvox work in the unicellular green alga Chlamydomonas and flanking sequences from Chlamydomonas work in Volvox, at least if a low expression rate is compensated by an enforced high gene dosage. This strategy might be useful for all investigators that intend to transform species in which genomic sequences are not available, but sequences from related organisms exist.

Keywords

Green algae Volvocales Recombinant DNA technology Transgenic expression Genetic transformation Paromomycin resistance 

Abbreviations

aphVIII

Aminoglycoside 3′-phosphotran sferase (VIII) gene

hsp70A

Heat shock protein 70A ge ne

rbcS2

Ribulose-1,5-bisphosphat-carboxylase small subunit gene 2 from Chlamydomonas reinhardtii

rbcS3

Ribulose-1,5-bisphosphat-carboxylase small subunit gene 3 from Volvox carteri

UTR

Untranslated region

Notes

Acknowledgements

We wish to thank Irina Sizova and Peter Hegemann for providing plasmid pSI103, Thomas Jakobiak and Rüdiger Schmitt for providing plasmid pPmr3, and Kordula Puls for expert technical assistance. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 521).

References

  1. Adams CR, Stamer KA, Miller JK, McNally JG, Kirk MM, Kirk DL (1990) Patterns of organellar and nuclear inheritance among progeny of two geographically isolated strains of Volvox carteri. Curr Genet 18:141–153CrossRefPubMedGoogle Scholar
  2. Buchheim MA, Chapman RL (1991) Phylogeny of the colonial green flagellates: a study of 18S and 26S rRNA sequence data. Biosystems 25:85–100CrossRefPubMedGoogle Scholar
  3. Buchheim MA, McAuley MA, Zimmer EA, Theriot EC, Chapman RL (1994) Multiple origins of colonial green flagellates from unicells: evidence from molecular and organismal characters. Mol Phylogenet Evol 3:322–343CrossRefPubMedGoogle Scholar
  4. Danilenko VN, Akopiants KE, Sizova IA, Michurina TA (1997) Determination of the nucleotide sequence and characterization of the novel aminoglycoside phosphotransferase aphVIII gene from the Streptomyces rimosus strain. Genetika 33:1478–1486PubMedGoogle Scholar
  5. Davies J, Wright GD (1997) Bacterial resistance to aminoglycoside antibiotics. Trends Microbiol 5:234–240CrossRefPubMedGoogle Scholar
  6. Debuchy R, Purton S, Rochaix JD (1989) The argininosuccinate lyase gene of Chlamydomonas reinhardtii: an important tool for nuclear transformation and for correlating the genetic and molecular maps of the ARG7 locus. EMBO J 8:2803–2809PubMedGoogle Scholar
  7. Hallmann A, Rappel A (1999) Genetic engineering of the multicellular green alga Volvox: a modified and multiplied bacterial antibiotic resistance gene as a dominant selectable marker. Plant J 17:99–109PubMedGoogle Scholar
  8. Hallmann A, Rappel A, Sumper M (1997) Gene replacement by homologous recombination in the multicellular green alga Volvox carteri. Proc Natl Acad Sci USA 94:7469–7474CrossRefPubMedGoogle Scholar
  9. Harris EH (1989) The Chlamydomonas sourcebook: a comprehensive guide to biology and laboratory use. Academic Press, San DiegoGoogle Scholar
  10. Jakobiak T, Mages W, Scharf B, Babinger P, Stark K, Schmitt R (2004) The bacterial paromomycin resistance gene, aphH, as a dominant selectable marker in Volvox carteri. Protist 155:381–393CrossRefPubMedGoogle Scholar
  11. Kindle KL (1990) High-frequency nuclear transformation of Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 87:1228–1232PubMedGoogle Scholar
  12. Kirk DL (1998) Volvox: Molecular-genetic origins of multicellularity and cellular differentiation. Cambridge University Press, CambridgeGoogle Scholar
  13. Lumbreras V, Stevens DL, Purton S (1998) Efficient foreign gene expression in Chlamydomonas reinhardtii mediated by an endogenous intron. Plant J 14:441–447CrossRefGoogle Scholar
  14. Provasoli L, Pintner IJ (1959) Artificial media for freshwater algae: problems and suggestions. In: Tyron CA, Hartman RT (eds) The Ecology of alga. Pymatuning laboratory of field biology, Special Publication no. 2, University of Pittsburgh, Pittsburgh, PA, pp 84–96Google Scholar
  15. Rausch H, Larsen N, Schmitt R (1989) Phylogenetic relationships of the green alga Volvox carteri deduced from small-subunit ribosomal RNA comparisons. J Mol Evol 29:255–265CrossRefPubMedGoogle Scholar
  16. Sanford JC, Smith FD, Russell JA (1993) Optimizing the biolistic process for different biological applications. Methods Enzymol 217:483–509PubMedGoogle Scholar
  17. Schiedlmeier B, Schmitt R, Müller W, Kirk MM, Gruber H, Mages W, Kirk DL (1994) Nuclear transformation of Volvox carteri. Proc Natl Acad Sci USA 91:5080–5084PubMedGoogle Scholar
  18. Schmitt R, Fabry S, Kirk DL (1992) In search of molecular origins of cellular differentiation in Volvox and its relatives. Int Rev Cytol 139:189–265PubMedGoogle Scholar
  19. Schroda M, Blöcker D, Beck CF (2000) The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. Plant J 21:121–131CrossRefPubMedGoogle Scholar
  20. Sizova I, Fuhrmann M, Hegemann P (2001) A Streptomyces rimosus aphVIII gene coding for a new type phosphotransferase provides stable antibiotic resistance to Chlamydomonas reinhardtii. Gene 277:221–229CrossRefPubMedGoogle Scholar
  21. Sodeinde OA, Kindle KL (1993) Homologous recombination in the nuclear genome of Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 90:9199–9203PubMedGoogle Scholar
  22. Starr RC, Jaenicke L (1974) Purification and characterization of the hormone initiating sexual morphogenesis in Volvox carteri f nagariensis Iyengar. Proc Natl Acad Sci USA 71:1050–1054PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Department of Cellular and Developmental Biology of PlantsUniversity of BielefeldBielefeldGermany

Personalised recommendations