A question that comes up from time to time is the use of co-infection with baculovirus vectors for multiple protein expression in insect cells. The need arises if you have produced a number of baculovirus vectors that each synthesise a component of a multi-subunit complex, such as an enzyme or a virus like particle. Intuitively, you know that it is probably necessary to add many virus particles per cell to achieve efficient infection with each recombinant. But how many viruses can you add to each cell and still get each one into every cell in equal numbers? (more…)
A question we are asked from time to time is to identify baculovirus transfer vectors
compatible with our flashBAC™ system. We obviously nominate our own extensive range that is available but frequently someone has an old plasmid from years ago that they want to reuse. (more…)
Variable expression of multi-component protein complexes is increasingly necessary as more sophisticated projects are attempted. But how can this be achieved? Most of our expression work is based on the baculovirus system, in particular our flashBAC™ vectors that permit easy recombinant virus generation in a one-step process. These insect-specific viruses offer an ideal platform for producing a wide range of recombinant proteins including nuclear, cytoplasmic, membrane-bound and secreted examples. (more…)
Between initial tests and larger scale up you may want to conduct intermediate scale recombinant protein production using baculovirus vectors. If you are working with several constructs then this can begin to get expensive of cell culture vessels such as the ubiquitous 125ml conical flasks, in which you can amplify 25-30mls of virus-infected cells. Space on shakers can also be at a premium in some labs. (more…)
Oxford Expression Technologies are the experts when it comes to the science of baculovirus. For almost 10 years now we have specialised in recombinant protein production in insect cells and the ‘Bac to the Future’ blog is the culmination of this extensive knowledge.
Written by our in-house scientists, articles cover every aspect of the baculovirus system – from its discovery and background, to practical applications in research and industry. We cover both the day-to-day challenges and more complex cases faced by ourselves and customers. You can also keep up to date on all the news, developments and promotional offers from the company.
If you have a question about any of the content featured, or simply want more information on a particular topic, then do not hesitate to get in touch with us directly at firstname.lastname@example.org. In addition, OET offers everything needed to aid your own insect virus research. As creators of the world renowned flashBACTM technology, our range of baculovirus expression vectors are designed for simple and rapid recombinant protein production. This is supported by an optimised range of transfer plasmids, transfection reagents, and cell culture media to ensure a powerful and versatile system. Browse the full collection of products at https://oetltd.com/shop/
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A question that often comes up from clients is what is the best signal peptide to use for baculovirus-expressed secreted recombinant proteins? While we have discussed the best flashBAC™ variant to use for the production of secreted proteins in a previous blog, we have never considered the role of the signal peptide. (more…)
There are many methods available for the titration of recombinant baculoviruses. One of the most convenient is the use of the quantitative polymerase chain reaction (QPCR), which assesses how much virus DNA is present in a freshly-amplified stock. This is compared with a standard curve derived from a control virus stock to give an infectious titre expressed as Q plaque forming units (pfu) per millilitre. OET Ltd market the baculoQUANT™ all in one virus extraction and titration kit for this purpose. (more…)
Sequence errors in your gene for expression Part II follows on from an earlier blog where we discussed a problem we were having producing a particular protein after synthesizing a gene based on very old sequence data. The baculovirus we made containing this synthetic gene produced very low levels of recombinant protein that were hardly visible after immunoblot analysis. This wasn’t a good start to a project where a purified protein was required for diagnostic purposes. (more…)
If you don’t achieve the levels of protein expression expected in your project, have you ever considered that there may be sequence errors in your gene? This applies to all expression systems, not just those based on baculoviruses. We should be clear that we are not talking about sequence errors in your gene after it has been synthesized by an out sourcing company. Quality control by such companies is such that it is extremely rare for there to be a problem. No, we are referring to data that you may have down loaded from online sources, principally GenBank. These databases host information from the very early days of sequencing, when deriving the sequence of an entire gene was a major achievement. (more…)
You may have had the experience! An expression project is going really well. You inserted your gene into a baculovirus vector (preferably one of our flashBAC™ range), amplified the virus, tested expression, obtained a rewarding blob of stained protein on a gel, went on to purify it – only to find and insoluble protein in your hands. Cue to retire to the bar for a consolation beer – in moderation of course! (more…)