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I'm sort of hating my master's thesis as I'm facing a setback after a setback... but even if I never would work as a biologist after this, I want to survive and finish my thesis and get a new boss. So... by far:1. I have managed to get the wanted protein fragment from the human cDNA library (the protein size is about 6000 bp, my fragment is from the c-terminal part and about 1000 bp)
2. I have cloned it into pET28 vector and expressed it in TOP10 and BL21(DE3) cells
I'm trying to purify the fragment now to sent it to the company... my goal is to get an antiserum against this protein's c-terminal part, which would stain tissues hopefully and which would at least change the world and give me a nobel price
ok, maybe I'm just trying to finish the thesis at first BUT, my thesis seem to be changing daily
My supervisor wants me to express this insert in mammalian cells now and I'm trying to find a vector, which would include a signal sequence (as I only have a protein fragment, not the entire protein) and a his tag. He told me to just check out websites and find one (ignore the fact that two days ago he told me to do something else ), but by far I haven't found any information about is expressing partial protein possible in any of the vectors. So I have two questions...1. Is this signal sequence normally included already in the vector?
2. Any suggestion about some good expression vector for a partial protein?
