protein cleavage

EK khatipovNO-SPAM at NO-SPAMuchicago.edu
Wed Oct 23 16:36:59 EST 2002


You don't provide enough details about your construct, so I assume you have
your tag in C-terminal fusion to your protein.If so, you most probably must
have left stop codon in the N-terminal frame. This, translation terminates
at this stop and you get only the protein without the tag. Could this be the
case? The only way to cure the problem is to redo the cloning making sure
there is no stop between fusion partners.
Emir

<j.ithri at caramail.com> wrote in message
news:20021023200925.26273.qmail at ww02.hostica.com...
>
> HI,
> i did a transient transfection to purify my protein with the complexe. but
when i did western to check for the transfection, i got a 32 kD band which
is my protein. but i was expecting a band at 52 kD (my protein with tag).
the only explanation i have is that my protein was cleaved. if so, that
means i can not start the purification.
> could someone help me to find the way that my protein would not be
cleaved.
>
> thanks
>
>
>
>
>
http://biowww.net/mynews/tree.php?group_name=bionet_molbio_methds-reagnts&be
gin=0





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