I hope I am posting in the right place.
I have some questions about the whole gene cloning affair in the bacteria host.
- I understand (theoretically) that plasmids in a host are replicated but do they always transcribe/translate (henceforth express) the encoded proteins?
- A cursory exam of plasmid maps show antibiotic resistance genes but do these genes have promoters if (1) is "yes"?
- The topic on gene expression control uses operon as example. Do these antibiotic genes get induced by anything or under any control?
- Can I simply clone a gene into a plasmid and get the host to show the new phenotype or do I need some sort of promoter to go with this gene (I am thinking along the line of PCR here)?
In the textbooks, the flow is always to obtain the gene of interest, clone it, and select for the plasmid bearing bacteria using the antibiotic markers or the expression of the gene. But is it that straightforward or are the details not mentioned?
I think there was this recent post about promoterless gene in here as well. And another one about two promoters. Both confuses me further.
Thank you to anyone who will patiently guide me through this.