The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri

von der Heyde EL, Klein B, Abram L, Hallmann A (2015)
BMC Biotechnology 15(1): 5.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
Background The multicellular green alga Volvox carteri represents an attractive model system to study various aspects of multicellularity like cellular differentiation, morphogenesis, epithelial folding and ECM biogenesis. However, functional and molecular analyses of such processes require a wide array of molecular tools for genetic engineering. So far there are only a limited number of molecular tools available in Volvox. Results Here, we show that the promoter of the V. carteri nitrate reductase gene (nitA) is a powerful molecular switch for induction of transgene expression. Strong expression is triggered by simply changing the nitrogen source from ammonium to nitrate. We also show that the luciferase (g-luc) gene from the marine copepod Gaussia princeps, which previously was engineered to match the codon usage of the unicellular alga Chlamydomonas reinhardtii, is a suitable reporter gene in V. carteri. Emitted light of the chemiluminescent reaction can be easily detected and quantified with a luminometer. Long-term stability of inducible expression of the chimeric nitA/g-luc transgenes after stable nuclear transformation was demonstrated by transcription analysis and bioluminescence assays. Conclusion Two novel molecular tools for genetic engineering of Volvox are now available: the nitrate-inducible nitA promoter of V. carteri and the codon-adapted luciferase reporter gene of G. princeps. These novel tools will be useful for future molecular research in V. carteri.
Co-transformation; Genetic engineering; Ammonium; Gaussia princeps luciferase gene; Green algae; Heterologous expression; Inducible promoter; Nitrate reductase; Nitrogen sources; Volvocine algae; Reporter genes; Streptomyces rimosus aphVIII gene
BMC Biotechnology
Open-Access-Publikationskosten wurden durch die Deutsche Forschungsgemeinschaft und die Universität Bielefeld gefördert.
Page URI


von der Heyde EL, Klein B, Abram L, Hallmann A. The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri. BMC Biotechnology. 2015;15(1): 5.
von der Heyde, E. L., Klein, B., Abram, L., & Hallmann, A. (2015). The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri. BMC Biotechnology, 15(1), 5. doi:10.1186/s12896-015-0122-3
von der Heyde, E. L., Klein, B., Abram, L., and Hallmann, A. (2015). The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri. BMC Biotechnology 15:5.
von der Heyde, E.L., et al., 2015. The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri. BMC Biotechnology, 15(1): 5.
E.L. von der Heyde, et al., “The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri”, BMC Biotechnology, vol. 15, 2015, : 5.
von der Heyde, E.L., Klein, B., Abram, L., Hallmann, A.: The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri. BMC Biotechnology. 15, : 5 (2015).
von der Heyde, Eva Laura, Klein, Benjamin, Abram, Lars, and Hallmann, Armin. “The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri”. BMC Biotechnology 15.1 (2015): 5.
Alle Dateien verfügbar unter der/den folgenden Lizenz(en):
Copyright Statement:
Dieses Objekt ist durch das Urheberrecht und/oder verwandte Schutzrechte geschützt. [...]
Access Level
OA Open Access
Zuletzt Hochgeladen
MD5 Prüfsumme

2 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Transgene Expression in Microalgae-From Tools to Applications.
Doron L, Segal N, Shapira M., Front Plant Sci 7(), 2016
PMID: 27148328
Algal Cell Factories: Approaches, Applications, and Potentials.
Fu W, Chaiboonchoe A, Khraiwesh B, Nelson DR, Al-Khairy D, Mystikou A, Alzahmi A, Salehi-Ashtiani K., Mar Drugs 14(12), 2016
PMID: 27983586

66 References

Daten bereitgestellt von Europe PubMed Central.

Kirk DL., 1998
Evolution of reproductive development in the volvocine algae.
Hallmann A., Sex. Plant Reprod. 24(2), 2010
PMID: 21174128
Nuclear transformation of Volvox carteri.
Schiedlmeier B, Schmitt R, Muller W, Kirk MM, Gruber H, Mages W, Kirk DL., Proc. Natl. Acad. Sci. U.S.A. 91(11), 1994
PMID: 8197189
Reporter genes and highly regulated promoters as tools for transformation experiments in Volvox carteri.
Hallmann A, Sumper M., Proc. Natl. Acad. Sci. U.S.A. 91(24), 1994
PMID: 7972102
The bacterial paromomycin resistance gene, aphH, as a dominant selectable marker in Volvox carteri.
Jakobiak T, Mages W, Scharf B, Babinger P, Stark K, Schmitt R., Protist 155(4), 2004
PMID: 15648719
Gene replacement by homologous recombination in the multicellular green alga Volvox carteri.
Hallmann A, Rappel A, Sumper M., Proc. Natl. Acad. Sci. U.S.A. 94(14), 1997
PMID: 9207115
regA, a Volvox gene that plays a central role in germ-soma differentiation, encodes a novel regulatory protein.
Kirk MM, Stark K, Miller SM, Muller W, Taillon BE, Gruber H, Schmitt R, Kirk DL., Development 126(4), 1999
PMID: 9895312
Genomic analysis of organismal complexity in the multicellular green alga Volvox carteri.
Prochnik SE, Umen J, Nedelcu AM, Hallmann A, Miller SM, Nishii I, Ferris P, Kuo A, Mitros T, Fritz-Laylin LK, Hellsten U, Chapman J, Simakov O, Rensing SA, Terry A, Pangilinan J, Kapitonov V, Jurka J, Salamov A, Shapiro H, Schmutz J, Grimwood J, Lindquist E, Lucas S, Grigoriev IV, Schmitt R, Kirk D, Rokhsar DS., Science 329(5988), 2010
PMID: 20616280
The molecular genetics of nitrate assimilation in fungi and plants.
Crawford NM, Arst HN Jr., Annu. Rev. Genet. 27(), 1993
PMID: 8122899
Nitrate: nutrient and signal for plant growth.
Crawford NM., Plant Cell 7(7), 1995
PMID: 7640524
Nitrate transporters in plants: structure, function and regulation.
Forde BG., Biochim. Biophys. Acta 1465(1-2), 2000
PMID: 10748256
Eukaryotic nitrate and nitrite transporters.
Galvan A, Fernandez E., Cell. Mol. Life Sci. 58(2), 2001
PMID: 11289304
Nitrate assimilation in Chlamydomonas.
Fernandez E, Galvan A., Eukaryotic Cell 7(4), 2008
PMID: 18310352
Root respiration associated with ammonium and nitrate absorption and assimilation by barley.
Bloom AJ, Sukrapanna SS, Warner RL., Plant Physiol. 99(4), 1992
PMID: 16669035
Genetic studies of nitrate assimilation in Aspergillus nidulans.
Cove DJ., Biological reviews of the Cambridge Philosophical Society. 54(3), 1979
PMID: IND80044864
PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family.
Quesada A, Krapp A, Trueman LJ, Daniel-Vedele F, Fernandez E, Forde BG, Caboche M., Plant Mol. Biol. 34(2), 1997
PMID: 9207842
Expression of nitrate assimilation related genes in Chlamydomonas reinhardtii.
Quesada A, Fernandez E., Plant Mol. Biol. 24(1), 1994
PMID: 8111016
Hsp70A and GlsA interact as partner chaperones to regulate asymmetric division in Volvox.
Cheng Q, Pappas V, Hallmann A, Miller SM., Dev. Biol. 286(2), 2005
PMID: 16168403

Cloning and characterization of new bioluminescent proteins
Szent-Gyorgyi C, Ballou BT, Dagmal E, Bryan B., 1999
Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo.
Tannous BA, Kim DE, Fernandez JL, Weissleder R, Breakefield XO., Mol. Ther. 11(3), 2005
PMID: 15727940
A multifunctional, synthetic Gaussia princeps luciferase reporter for live imaging of Candida albicans infections.
Enjalbert B, Rachini A, Vediyappan G, Pietrella D, Spaccapelo R, Vecchiarelli A, Brown AJ, d'Enfert C., Infect. Immun. 77(11), 2009
PMID: 19687206
Optimisation of bioluminescent reporters for use with mycobacteria.
Andreu N, Zelmer A, Fletcher T, Elkington PT, Ward TH, Ripoll J, Parish T, Bancroft GJ, Schaible U, Robertson BD, Wiles S., PLoS ONE 5(5), 2010
PMID: 20520722
Alternative luciferase for monitoring bacterial cells under adverse conditions.
Wiles S, Ferguson K, Stefanidou M, Young DB, Robertson BD., Appl. Environ. Microbiol. 71(7), 2005
PMID: 16000745
Biophysical characterization of highly active recombinant Gaussia luciferase expressed in Escherichia coli.
Rathnayaka T, Tawa M, Sohya S, Yohda M, Kuroda Y., Biochim. Biophys. Acta 1804(9), 2010
PMID: 20452471
Stable nuclear transformation of Gonium pectorale.
Lerche K, Hallmann A., BMC Biotechnol. 9(), 2009
PMID: 19591675
Stable nuclear transformation of Eudorina elegans.
Lerche K, Hallmann A., BMC Biotechnol. 13(), 2013
PMID: 23402598
Stable nuclear transformation of Pandorina morum.
Lerche K, Hallmann A., BMC Biotechnol. 14(), 2014
PMID: 25031031
Triassic origin and early radiation of multicellular volvocine algae.
Herron MD, Hackett JD, Aylward FO, Michod RE., Proc. Natl. Acad. Sci. U.S.A. 106(9), 2009
PMID: 19223580
A secreted luciferase for ex vivo monitoring of in vivo processes.
Wurdinger T, Badr C, Pike L, de Kleine R, Weissleder R, Breakefield XO, Tannous BA., Nat. Methods 5(2), 2008
PMID: 18204457
Chemically inducible promoters in transgenic plants
Gatz C., 2014
Ecdysone agonist inducible transcription in transgenic tobacco plants.
Martinez A, Sparks C, Hart CA, Thompson J, Jepson I., Plant J. 19(1), 1999
PMID: 10417731
Expression profiling of the maize flavonoid pathway genes controlled by estradiol-inducible transcription factors CRC and P.
Bruce W, Folkerts O, Garnaat C, Crasta O, Roth B, Bowen B., Plant Cell 12(1), 2000
PMID: 10634908
Structure, reproduction and differentiation in Volvox carteri f. nagariensis Iyengar, strains HK 9 & 10
Starr RC., 1969
Control of differentiation in Volvox
Starr RC., 1970
Artificial media for fresh-water algae: problems and suggestions
Provasoli L, Pintner IJ., 1959

Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction.
Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden TL., BMC Bioinformatics 13(), 2012
PMID: 22708584
A simple and rapid method for the preparation of plant genomic DNA for PCR analysis.
Edwards K, Johnstone C, Thompson C., Nucleic Acids Res. 19(6), 1991
PMID: 2030957
Molecular toolbox for studying diatom biology in Phaeodactylum tricornutum.
Siaut M, Heijde M, Mangogna M, Montsant A, Coesel S, Allen A, Manfredonia A, Falciatore A, Bowler C., Gene 406(1-2), 2007
PMID: 17658702
Enzymes involved in anaerobic respiration appear to play a role in Actinobacillus pleuropneumoniae virulence.
Jacobsen I, Hennig-Pauka I, Baltes N, Trost M, Gerlach GF., Infect. Immun. 73(1), 2005
PMID: 15618158
New spectrophotometric equations for determining chlorophyll a, b, c and c in higher plants, algae and natural phytoplankton
Jeffrey SW, Humphrey GF., 1975


Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®


PMID: 25888095
PubMed | Europe PMC

Suchen in

Google Scholar