Expression Vector vs Cloning Vector

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testsubjectzz
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Expression Vector vs Cloning Vector

Post by testsubjectzz » Sat Sep 25, 2010 1:32 pm

I have this question how come with cloning why can we not clone directly into the expression vector why is it done in 2 stages?

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JackBean
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Post by JackBean » Sat Sep 25, 2010 2:52 pm

Sure you can and sometimes you even do, like e.g. if you're expressing in bacteria or have vector working both in bacteria and yeast (like pPICZ and pGAPZ).

However, the point of using two vectors is such, that you can use your protein for plenty of applications (e.g. expression, GFP fusion, crystalization etc.) and if you had to get it always from either gDNA or mRNA would be quite hard so you better clone it into one (clone) vector, which is high copy, but doesn't bother the bacteria with expression of your protein and thus you can always obtain high amount of your gene.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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