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
 
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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.
Stichworte
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
Erscheinungsjahr
2015
Zeitschriftentitel
BMC Biotechnology
Band
15
Ausgabe
1
Art.-Nr.
5
ISSN
1472-6750
Finanzierungs-Informationen
Open-Access-Publikationskosten wurden durch die Deutsche Forschungsgemeinschaft und die Universität Bielefeld gefördert.
Page URI
https://pub.uni-bielefeld.de/record/2722409

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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, Eva Laura, Klein, Benjamin, Abram, Lars, and Hallmann, Armin. 2015. “The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri”. BMC Biotechnology 15 (1): 5.
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.
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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
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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

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