I would like to know whether its possible to clone and overexpress a large protein of the size of approx 450-500 kDa without truncating any part of the protein?
so you would be looking to clone 14-16Kb of DNA. I think that would be possible but you would have to be careful about the stability of the DNA in E coli. I think your problem would be expression. Theres no way that it would express in E coli, Im not sure if you would get so much DNA in a Bacmid for insect cell expression which leaves you with mammalian expression.
You might spend 6 months cloning it to find out it wont express
good luck though!
Mark
You COULD consider yeast as the expression system (yeast does express so big and even bigger proteins; http://www.biomedcentral.com/1471-2148/6/61/), but the recommendations above seems more plausible.
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