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Studying genes at the Trancription& Translation level
I working on a strain that doesn't require a substrate to express the enzyme whereas other strian among the smae species require to. So, I am lil bit confused on how do I go about the whole thing?
Do I have to get the total RNA, do a real time PCR etc;??
Re: Studying genes at the Trancription& Translation level
You likely want a quantitative Real-time-RT-PCR (one-step rev-trancription-PCR is pretty nice) and maybe an ELISA or quantitative WB.
Another option for protein-of-interest quantitation, is either immuno-affinity or tag-specific purification, then a simple BCA protein assay of purified protein (with a SDS-gel for purity confirmation, gel quantitation w/ a standard and appropriate software is also possible).
Or if the enzyme activity can be measured, you can measure enzyme activity relative to amount of mRNA per ug sample/#cells (or whatever quantity).
|genes , level , studying , trancriptionand , translation|
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