In summary, our research group works on proteins contributing to cell polarity, on proteins acting on the cell surface during male-female gamete interactions, and on the role of Argonaute proteins and small RNAs in plant reproduction.
To address questions related to our research topics we establish protocols to isolate living gametes from different flowering plant species. We perform transcriptomics and proteomics to identify candidate genes and proteins that are subsequently tested in functional studies.
Currently, we are investigating the molecular function of a small protein family that participates in the establishment and maintenance of cell polarity by characterizing specific interaction partners. Furthermore, we study (small) RNAs and proteins putatively involved in egg cell specification, and characterize cell surface proteins which are involved in gamete adhesion, gamete activation, or fusion.
Methods
We are using a wide range of basic and advanced methods related to (plant) molecular biology, molecular genetics, cell biology and protein biochemistry.
- DNA work (DNA isolation, conventional & Gateway?-based cloning, PCR, real-time quantitative PCR, PCR-mediated mutagenesis, Overlap extension PCR cloning, RNAi, amiRNA, CRISPR/Cas9, Southern Blot, Southwestern Blot, cDNA libraries, etc.)
- RNA work (total RNA and mRNA isolation, small RNA isolation, cDNA synthesis, RT-PCR, In vitro-transcription, DIG labeling, In situ hybridization, Northern Blot, etc.)
- Protein work (protein extraction, PAGE, 2D-PAGE, Western Blot, protein staining, immunoprecipitation, protein phosphorylation, recombinant protein expression, affinity purification, Size Exclusion Chromatography, ?kta pure system, etc.)
- Interaction studies (Y1H and Y2H screens, EMSA, Co-immunoprecipitation, etc.)
- Microscopy (fluorescence microscopy, DIC microscopy, Confocal Laser Scanning Microscopy, Spinning-disc live-cell imaging, immunofluorescence, etc.)
- Transformation of bacteria and yeast
- Plant transformation (biolistic, Agrobacterium-mediated) and genetics
- Plant cell culture (Arabidopsis, wheat, maize)
- Micromanipulation (single cell isolation)
- Expression studies (Microarrays, RNAseq, RT-PCR)