peter.cherepanov at uz.kuleuven.ac.be
Sat Aug 3 05:19:28 EST 2002
>I wanted to know what does rapid turn over mean? And also wanted to have a
>protocol for the purification of HBx. I am basically an inorganic chemist.
1) turover - reflects the rate of the protein's degradation in the host cell
(by host cell proteases, I suppose). You can measure it by a pulse-chase:
labeling your infected cells with a radioactive amino acid for some time and
then adding an excess of cold amino acid (chasing). Then you isolate the
protein and measure the specific radioactivity of it (how fast it drops) -
it should give you the idea about turover.
2) purification from E. coli - this you should do according to the existing
manuals. Check the Novagen's and Qiagen's web sites. Both have extensive
manuals for His-tagged protein purification from bacteria.
I think, it would be great if you could find somewhere a protein
biochemistry lab, it will surely help and save you a lot of time.
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