IPTG and gene expression in pBluescript vector

[biozita]

IPTG and gene expression in pBluescript vector

• Quote

Post by biozita » Thu Aug 09, 2007 5:25 pm

Hi

I have cloned a kanamycin gene in the lacZ of pBluescript vector. However I'm not sure if I need to put IPTG during the expression time after transformation.
Anybody knows anything about it?

thanks

[blcr11]

• Quote

Post by blcr11 » Thu Aug 09, 2007 8:44 pm

It's not immediately obvious to me why you would want such a construct, but I'm sure there must be a good reason. If you really are trying to express the beta-gal-kanamycin fusion protein then you will need to induce with IPTG, just as you did when you did the blue-white screening. I'm assuming you've done the blue-white screening, since you say you've cloned the gene already. Just for expression of the fusion protein, you don't need to add X-gal, but you still need the IPTG.