I'm interested in using E. Coli to express a protein of interest.
I'm currently working on designing the plasmid. I know that the following are imperative for my plasmid:
1) Promoter, along with other host-specific elements for expression
2) My cDNA with a C-Terminal His Tag
3) Antibiotic Resistence Gene
I started looking online at expression plasmids, for instance: pTrisHis by LifeTechnologies. With this, I would simply need to cut my cDNA and the plasmid with the same RE, add them together, and ligate.
However - the cost made me rethink my strategy. Is it possible to isolate plasmids from your E. Coli strain (their genome is sequenced, I presume), and design an expression plasmid based off of that? Their plasmids should already have the Promoter and other expression elements - would I just need use a restriction map and add my cDNA and Antibiotic gene?