Lerche, Kai; Hallmann, Armin: Stable nuclear transformation of Gonium pectorale. In: BMC Biotechnology. Jg.9 H. 1. 2009
Inhalt
- Background
- Results
- Phylogenetic analysis of utilized Gonium pectorale strains
- Preliminary transformation studies using aphVIII-based selectable marker plasmids
- Recovery of transformants after biolistic transformation
- Paromomycin resistance of transformants
- Stable integration of plasmid DNA into the genome of transformants
- Detection of AphVIII protein in transformants
- Stable co-transformation of unselectable genes
- Transcription of co-transformed, unselectable genes
- Analysis of heterologous protein expression in co- transformants
- Long term stability of DNA integration and gene expression
- Discussion
- Conclusion
- Methods
- Strains and culture conditions
- Transformation vectors
- Preparation of plasmid DNA
- Coating of microprojectiles
- Determination of cell concentration
- Stable nuclear transformation by particle gun
- Re-isolation of transformants
- Paromomycin-resistance assay
- Primer design
- Isolation of genomic DNA
- Genomic PCR
- Southern blotting
- Isolation of total RNA
- Reverse Transcription (RT)-PCR
- Western blot analysis
- Luciferase assays
- Phylogenetic analysis
- GenBank accession numbers
- Abbreviations
- Authors' contributions
- Additional material
- Acknowledgements
- References