Sometimes less is more! Return to simpler times for an inexpensive way to get your gene of interest. In the fifth addition to our Vectors and Viruses Spotlight Series, we'll introduce you to ORF, a team player that can help you clone your gene into other vectors. Learn the advantages of choosing to use ORFs in your experiments in the article below!
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"Hello, you can call me Ophelia. I’m a simple, down-to-earth sort of vector. I don’t have any fancy promoters or packaging signals. But, I’m a real team player, and can help clone your gene into other vectors."
For doing your own cloning, it’s best to work from a simple non-viral plasmid carrying your gene of interest. These are called Open Reading Frame (ORF) vectors, as they contain only the translated region of the gene without the 5’ or 3’ untranslated regions. They also lack extraneous elements such as promoters or reporters.
An ideal ORF vector is one that has a variety of restriction enzyme sites available on either side of the insert. This allows it to be easily cloned into an assortment of new vectors using traditional ligation. However, even without convenient restriction sites, an ORF can be used for ligation-free cloning by amplifying it via PCR.
There are a few advantages of working from an ORF vector instead of a synthesized gene. First, an ORF vector can be easily amplified via bacterial transformation, so there is no risk of running out of DNA. Second, large genes may be less expensive to purchase as an ORF vector than they would be to synthesize. Finally, it can be difficult to synthesize sequences that contain repeated elements or have a high GC-content; for these sequences, it would be easier to use an ORF vector.
abm offers a complete genome-wide collection of human, mouse, and rat ORFs, ready to use in your cloning project. With both the 5'UTR and the 3'UTR removed, ORFs allow for quick and reliable protein expression when inserted into a compatible expression vector. They can also be used as a template for positive control in PCR amplification experiments.
Don't know which viral vector to choose?
You know you want to express a gene, but when it comes time to choose an expression vector, your choices can seem overwhelming. Do you go viral or non-viral? Which virus should you use? Or maybe it's not a protein you need to express, but an RNA species?
Try using our Vector Selection Tool. Cut through the fog with this Vector Selection Tool. Simple, clear recommendations for every application.
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