Protein production success!

Hurry up and wait. That's what we do in lab. Some weeks are pronounced with rapid developments and others, not so much... :)

I am happy to report that our team has already been successful in cloning our antiviral gene into a bacterial expression vector and we have been able to induce protein expression. The figure below is a Western blot of lysates from uninduced (---) and induced (ADACO) E. coli that contain our expression vector. We used an antibody that recognizes the Apaf1 domain of ADACO to reveal our protein on a nitrocellulose membrane.

 

Now that we can induce expression of our protein in E. coli, we should be able to purify it by taking advantage of a GST tag incorporated at its N-terminus. Once we purify the protein, we will determine if its antiviral activity is still effective against extracellular virus.

We hope that this protein will have both antibody-mediated neutralizing and apoptosis-inducing activities. This is the 1-2 punch that will protect cell cultures from infection without selecting for adaptive mutations. Keep your fingers crossed.