I have cloned my target gene (From plant) in pET32a but i cant transform it in BL21 (DE3) cells. I tried many times its not getting transformed. Even i tried along with pLYS S but i am not getting.. In this case, can i use XL1 Blue cells to check protein induction using IPTG?
Since genes cloned in pET vectors are under the control of a T7 promoter, it is unlikely that you will get satisfactory expression in a strain like XL1 Blue that doesn't carry the gene for T7 RNA polymerase. It sounds like your gene expresses a toxic protein. I would recommend using a strain that affords a tight control of T7 polymerase expression, like BL21AI (L-arabinose-inducible), available from Invitrogen
Thank you sir, But long back i have successfully transformed this same construct to BL21(DE3) and got expression, that i have checked it in SDS PAGE. Now i wanted to repeat this, But i am not getting colonies after transformation,
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