This is a modified pMal vector. We put the MBP protein in the C terminus, rather than the N terminus.
I first tried to express in BL21(DE3)STAR (as always). induced with IPTG (tried 0.3mM, 0.5mM, 1mM) when OD600 reached 0.5-0.6; expressed in 37C (2-4h), 25C,18C,14C (Overnight),
I took out samples before induction and after expression, measured the OD (to make sure I load the similar numbers of cells to the SDS-PAGE). Boiled in sample loading buffer , then run the gel.
I didn't see any difference between the un-induced and the induced.
I tried to purify some proteins, and I did get the band I wanted.
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#2
mdfenko
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Posted 03 August 2010 - 06:51 AM
if you got your protein then it was autoinduced.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#3
rosepolaris
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Posted 05 August 2010 - 06:02 AM
Thanks!
The yield was extremely low, and I need a lot of proteins to do crystallization..... Oh my!
I was wordering if there's a specific cell strain I need to try....
The yield was extremely low, and I need a lot of proteins to do crystallization..... Oh my!
I was wordering if there's a specific cell strain I need to try....
mdfenko, on 03 August 2010 - 06:51 AM, said:
if you got your protein then it was autoinduced.
