|High recombinant protein expression level using recombinant baculoviruses|
It is now routinely possible to obtain similar protein expression levels using recombinant baculoviruses as those typically observed in E.coli. Using a baculovirus system, recombinant protein expression levels often reach several percent of the total cellular protein. With the exception of very large or toxic proteins, expression levels are high enough that recombinant proteins can be seen as prominent bands on Coomasie-stained gels of cell extracts. As a general rule, it is not even necessary to utilize Western blotting for the initial recombinant protein expression analysis when using recombinant baculoviruses.
The goal of this tutorial is to help you to achieve the maximum expression level of your protein of interest in the baculovirus system. Many researchers who experience low levels of recombinant protein expression believe that it is an inherent characteristic of their protein of interest, while in reality they have failed to realize the potential of the baculovirus system. Overall, the baculovirus system is believed to be cumbersome, and unlike mammalian or E.coli systems few labs are proficient in using it.
AB Vector believes that this is not an inherent drawback of the system, but the lack of appropriate guidance. The few high quality guides written by "the founding fathers" of the system are outdated and hard to obtain. Technical documentation accompanying vectors, insect media and cells is often lengthy, complex and targeted to a particular product while missing important details and lacking overall perspective. It is written by professional writers of technical documentation rather than by the hands-on researchers who developed the products.
Having up to 20 years of hands-on experience with developing baculovirus products, we understand pitfalls of the system and have captured our experiences in this simple tutorial. Contact us at firstname.lastname@example.org if you have any questions, suggestions or comments. We would like to incorporate your feedback into this tutorial to help others to become successful users of the baculovirus system.