membrane protein expression

silkebd at gmx.de silkebd at gmx.de
Fri Nov 8 09:09:00 EST 2002


Hi,
I am trying to express in E. coli four different membrane proteins (ABC transporter) in either a pET vector with an N-terminal 6x his-tag or an pASK-IBA vector (tet promotor) with N- or C-terminal strep-tag, respectively. I tried different concentrations of inductor, different times of expression, different temperatures... Under all condition the proteins seem to be toxic (decrease in OD), and I cannot detect anything of the right size after fractionation and column purifcation or just fractionation western blotting, antibody detection.
Has anyone an idea how I can get my proteins?

Thanks for your advice!
punzel

http://biowww.net/mynews/tree.php?group_name=bionet_molbio_methds-reagnts&begin=0



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