Hi all, is it normal after conduct an E.coli transformation, there were colonies that having low producing protein target and the other having higher producing? What factor(s) that affect this result? Thank you.
Hello,
But once you collected different colonies, what did you do? A liquid culture?
If so, have you done a count to quantify the number of bacterial cells in your culture or an optical density measurement at 600 nm, to see if the cultures are identical in terms of cell number.
Maybe you just didn't have the same amount of cells after culture.
I have seen this often but don't have an explanation. We would usually screen a dozen or so independent transformants for expression levels and work with the one or two best ones.
In my opinion, It is the toxicity of the target protein towards the expression host that affects its growth.
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