Hellow!I am overexpressing GST-fused protein in PET-41a vector in BL21(pLysS)host strain now.I have totally five proteins for overexpression.But only two get overexpressed.I tried the different incubation temperature and time,and with detergent addition on the other three,but failed.I have tested the plasmid stability,and find it's stable.So what's going on in my work?My proteins are big,from 60kd to 90kd.Could you give me some advice about it?Thank you very much!!!!!
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