von der Heyde, Eva Laura ; Klein, Benjamin ; Abram, Lars; Hallmann, Armin: The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri. In: BMC Biotechnology. Jg.15 H. 1. 2015
Inhalt
- Results
- Chimeric selectable marker and reporter genes for transformation
- Generation of transgenic Volvox strains using the aphVIII selectable marker
- Genomic integration of chimeric gene constructs
- Expression of the g-luc reporter gene in transformants
- Long term stability of DNA integration and gene expression
- Inducibility of the nitA promoter-driven g-luc reporter gene in transformants
- mRNA and protein expression kinetics after induction of the nitA promoter-driven chimeric reporter gene
- Discussion
- Conclusion
- Methods
- Strains and culture conditions
- Transformation vectors
- Coating of microprojectiles
- Stable nuclear transformation by particle bombardment
- Paromomycin-resistance assay
- Primer design
- Isolation of genomic DNA
- Genomic PCR
- Isolation of total RNA
- Quantitative Real-Time RT-PCR
- Luciferase assays
- Chlorophyll determination
- mRNA and protein expression kinetics
- Abbreviations
- Competing interests
- Authors’ contributions
- Acknowledgements
- References