bicistronic retroviral vector - Two both gene mammalian expression (Nov/04/2005 )
I would like transfect 2 genes in the same retroviral vector. These 2 genes are necessary to immortalize eukaryotic cells.
Someone can help me for the choice of this one.
You could have either two separate promoters (one being in the LTR and the other internal) or utilize an IRES element. IN either case, it is a good idea to have a selectable markers. If you express a protiens from an IRES be reay to have a lower level of expression compared to the first cistron.
i don't know any retroviral vector that can do your job. They usuammy have a selectable marker under ltr and an internal promoter.
In other cases, selectable marker is under control of a promoter and you can clone a cDNA under an other one.
Hnce you would have to build your own vector. You can check for possibilities of digesting IRES sequence from an other one and clone it to your vector. With good ligations, this may be done relative quickly.