Can I clone and express a yeast gene of beta galactosidase into a bacterial host E. coli BL21? Will it effect the functionality of the protein?
Hey if you are using shuttle vector you can do it. If you mean with yeast
Saccharomyces cerevisiae, you can find easily shuttle vectors.
Why not, there are many examples of human gene expression in bacteria, so of course it can be done.
Dear Deepika,
Yes, you can. However, you must make sure of certain things. In E.coli, you have to have an idea of what will you be attempting, intracellular expression or soluble expression ?! If you are attempting soluble expression, what signal sequence would you be adding to the gene sequence?! The functionality of your protein could be assessed only after an experimental trial post cloning; In majority of the cases your protein would remain functional (you may screen it using a plate based or colorimetric assay). In order to maximize expression and product recovery, you will have to think of the methods, which I have listed above. WHy don't you attempt expression in PIchia pastoris? Just a suggestion!!
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