Hi,
I am actually just starting a project to transfect chlorella too. I also don't have a fixed protocol to perform this, although I have experience with electroporating other cell types, from e-coli to human cells. The main problem with chlorella seems to be that its cells are rather small as far as eukaryotic cells are concerned, and they have a thick cell wall. So first one needs to digest their cell wall away before electroporation. Alternatively a gene gun can also be used with which transfection is possible even without digestion. I understand the latter is the weapon of choice usually when transfecting chlorella.
Finding the genomic integration site will be another problem, but this might be done with a simple PCR and sanger sequencing of the product, using a primer within the plasmid and a bunch of random primers. This may or may not work, if it doesn't then probably some high throughput sequencing will need to be done, which will quickly get rather expensive if you want to test multiple single-cell colonies.
Either way, I'd be interested in staying in touch and developing a good protocol for this as I'm in need of it too.
Cheers,
Mate
On Wednesday, 11 July 2018 11:49:00 UTC+1, najib abdellaoui wrote:
hello everyonei am currently working on a project for production of vaccine based on stable transformation of chlorella vulgaris. the plasmid size is around 6 Kb and i wanted to transformed into chlorella vulgaris using electroporation to get a stable expression of my protein of interest and also i wanna analyze the site of integration into genome. we have bio-rad gene pulser II in our lab.so can someone help me for the design of protocol for the transformation.thanks in advance.
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