I have cloned a mutant human polb sequence in a pET28a vector. I have sequenced the clone, the sequence is in reading frame. When I try to expresses the protein in BL21 with IPTG, get no product. I have tried for several time, also in different concentration of IPTG, time period and temperature. Please help me.
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#2
bob1
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Posted 10 April 2017 - 07:53 AM
Which fractions have you solubilised and done westerns for?
#3
labtastic
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Posted 10 April 2017 - 09:27 AM
And do know this protein should express in E.coli (from literature, previous lab members, etc)? Sometimes proteins just don't express in Ecoli.
Edited by labtastic, 10 April 2017 - 09:31 AM.
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