I want to express bacterial gene into eukaryotic cells, yeast to be precise. Does someone know some article about this?
Hi,
There is a whole book about gene expression and protein purification in Pichia pastoris yeast. Please access it here
If you have further questions, please let me know.
Girish
http://www.springer.com/us/book/9781588294296
This is not a problem. All you need to do is clone your bacterial coding region into a eukaryotic plasmid vector that contains a good promoter and a poly-adenylation site at the 3' end. There are many examples of bacterial genes being used in eukaryotes, LacZ, neomycin resistance, tetracycline resistance, etc.
There should be plenty of information. on the web. This is a standard technique we teach undergrad students. You clone the gene and put it in a plasmid that can be used in both E. coli and yeast. You first transfect E. coli and grow a culture to obtain a large number of plasmids. Purify the plasmids and transfect them into yeast with LiAc and heat shock. You use lacZ or antibiotics resistance to select succesfully transfected E. coli. Then use use an auxotrophic yeast strain to select the transfected yeasts.
Thanks a lot Girish , Gregory and Jonas for your help.
I know the book Girish spoke about. I looked extensively in the web and I found a lot about what you said, Gregory. I think I didn't explained myself plainly. I want to build a system to knockout a gene in the yeast, S. Pombe for instance, replace it by a bacterial gene (ortholog) and see it there is expression of this gene.
I am grateful to you for answering me.
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