I am very new to the concept of gene cloning and transformation. I am trying transfer the arenite oxidase gene cluster (9kb in length) to E. coli strain C600. I want the E. coli strain to express this gene in order to oxidize arenite for bioremediation of groundwater. I am just realizing how complicated it really is to select the right expression vector. I have not studies this topic at all at school (still in high school); I am just studying this topic on my own. I am worried that my 9kb sequence could be too big for the plasmid vector or the bacterium. I have been trying to find the right plasmid expression vector for days, and it seems like everyone I find isn't quite right.
I have been kind of vague, so here is the whole story: I am in possession of a gram-negative bacteria that can oxidize arsenite into arsenite very efficiently. The gene cluster that is responsible for this is Aro, which is 9 kilobases in length, and I am looking to use a heat shock method to transfer that gene cluster into a specific bacterium (I found that E. coli C600 might be a worthwhile candidate). I have everything I need for the extraction of the DNA and PCR, but I am just trying to find a plasmid expression vector that I can insert into E. coli C600. Originally, I looked at pUC19, but then I found out it is only a cloning vector and not an expression vector. Then I looked at the pET system, but there are so many selections; I cannot seem to find the right one.
Are there any suggestions for what vector I should use or any guidance on how to find one?
