Lerche, Kai; Hallmann, Armin: Stable nuclear transformation of Pandorina morum. In: BMC Biotechnology. Jg.14 H. 1. 2014
Inhalt
- Background
- Results
- Antibiotic tolerance of wild-type Pandorina algae
- Transformation experiments using aphVIII as a selectable marker
- Paromomycin resistance of transformants
- Stable genomic integration and expression of the artificial aphVIII gene
- Calculation of transformation frequency
- Plasmids for co-transformation with non-selectable reporter gene constructs
- Stable integration of non-selectable reporter gene constructs into the genome
- Functional expression of co-transformed reporter genes
- Long term stability of DNA integration and gene expression
- Discussion
- Antibiotics resistance
- Transformation frequency
- Co-transformation rate
- Expression of heterologous genes using heterologous promoters
- Conclusion
- Methods
- Strains and culture conditions
- Transformation vectors
- Preparation of plasmid DNA
- Coating of microprojectiles
- Determination of cell concentration
- Stable nuclear transformation by particle bombardment
- Re-isolation of transformants
- Paromomycin-resistance assay
- Primer design
- Isolation of genomic DNA
- Genomic PCR
- Isolation of total RNA
- Reverse Transcription (RT)-PCR
- Induction of luciferase activity
- Luciferase assays
- In-gel activity assay
- Additional file
- Abbreviations
- Competing interests
- Authors’ contributions
- Acknowledgements
- References