2. Stock, Raum Nr.218
5. Stock, Raum 5X17
4 sterile benches, growth rooms
Our New Bench: Mission Impossible...
Dr. Kenneth W Berendzen
|Caterina Brancato|| |
Service provided by Anja and Caterina
Transformation Unit Protocols
- Cell Culture 01.docCell Culture55 K
- BY-2-protopl.docBY-2 Tobacco Cell Suspensions Protoplasts54 K
- PEG Cell Susp Transf Protopl2017.pdf0.9 M
- PotatoStabile.docTransformation protocol for potato52 K
- Transformation protocol for tobacco68 K
- Protoplasts isolation from leaves of Arabidopsis thaliana3.3 M
- Protocol for tomato stable transformationProtocol for tomato stable transformation. 95 K
|1. The unit offers the service of Agrobacterium-mediated transformation of potato, tobacco and tomato.|
|2. Regenaration and cultivation of transformants.|
|3. PEG-mediated transformations of protoplasts from cell cultures or plants-mesophyl (Arabidopsis or tobacco).|
|4. Productions of cell cultures (wild type).|
Fig. Z-stack image of protoplasts liberated from Arabidopsis thaliana leaf tissue.
Fig. Calculation of the protoplat transformation efficiency by flow cytometric anaylsis (FCA).
Protoplasts isolated from root cell culture are transformed with a 35S::GFP construct (12kb) as a routine control. As an alternative to calculating the transformation efficiency, we have used FCA as a means for detecting cells expressing GFP. The negative population is marked in black. Cells that are strongly expressing GFP have a fluoresence signal that is well above that of the total plant cell population‘s green autofluoresence (cells marked in dark green); cells marked in light green are those with weaker GFP fluoresence emission. In this example 23% of the cells were transformed with GFP, and 50% of those are strongly expressing GFP.
Albert I, Böhm H, Albert M, Feiler CE, Imkampe J, Wallmeroth N, Brancato C, Raaymakers TM, Oome S, Zhang H, Krol E, Grefen C, Gust A, Chai J, Hedrich R, Van den Ackerveken G, Nürnberger T (2015). An RLP23-SOBIR1-BAK1 complex mediates NLP-triggered immunity. Nature Plants, 15140, doi: 10.1038/NPLANTS.2015.140.
J. Wittmann, C. Brancato, K. W. Berendzen and B. Dreiseikelmann (2015).
Development of a tomato plant resistant to Clavibacter michiganensis using the endolysin gene of bacteriophage CMP1 as a transgene.