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polb cloned in pET not expressed

Protein Expression

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#1 Rana2017

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Posted 08 April 2017 - 07:09 AM

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.



#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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