Hi, just wondering... is it possible to induce overexpression of my protein cloned into pcDNA3 vector in bacterial cells? In teory it has T7 promoter but this aproach was not working in my hands? Any experience?

I have used KRX cells with T7 RNA polymerase controled by rhamnose promoter but using rhamnose as inductor did not work..., but I have only little experience so maybe just some stupid mistake

thanks

---

Which bacterial cells do you use?

---

KRX but if it would help, then I can change them

---

we use BL21(DE3)RIL cells for protein production. And I have seen many papers where these is used for protein production. its from stratagene.
I am not an expert in this field to say that this is the best or not.

---

I could try BL21 strain, they also have T7 RNA polymerase but under control of lac promoter... but I am afraid that my problem is not in bacterial strain but more likely in plasmid itself. Anyway I can still clone it into different plasmid... but I was just curios if it is possible with this vector.

---