Dear Aime,

I cannot answer your question with absolute certainty, as I did not test this particular situation. But we have tested the EC1.2 promoter previously in somatic cells (which is the equivalent to your protoplasts, just without the process of protoplasting), and it was not active at all. So most likely, you will not see any activity, irrespective of whether your system works or not (in stable transformations).

Otherwise, we have tested the EC promoter fragment in different vector architectures. And results are somewhat odd: We obtained very good results in some experiments (see for example a small publication from my lab including these results, here as preprint https://www.biorxiv.org/content/10.1101/393439v1). In other vector architectures, it did not work at all. And I have also heard from someone else that observed similar things. So if your system was not previously tested, you will have to see after stable transformation.

Therefore, EC1.2 is not my favorite promoter, although it can work nicely and basically eliminates chimeric plants.

We routinely use the RPS5a promoter, which is really giving great results.

Good luck for your experiments!

Bests,

Johannes

Dear Johannes Stuttmann ,

Thank you for your answer! And sorry for the lateness of mine. I have my T1 generation now and fortunately the system works really good for most of my constructs. I have a lot of homozygous plants in this first generation, so I'm really happy :)

Best,

Aime

Hi, if you say many, can you give a percentage? Always curious.

And great to hear that everything works fine, all the best for your experiments