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Baculovirus Expression Vector Systems


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An animation of the basic theory behind and workflow of Baculovirus Expression Vector Systems for recombinant protein expression in insect cells.

Published in: Science
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Baculovirus Expression Vector Systems

  1. 1. BACULOVIRUSES are natural viruses that infect insects in the wild But we can “unnaturalize” them to turn them into custom protein-making factories arly in the infection, the cells make and secrete budded viru They use this to jump into other cells in the same insectBut the insect gets really sick and can’t support the virusThey have circular, double-stranded DNA genomesTheir goal is to get into insect cells to make more of them. And they can do this a couple of ways
  2. 2. starts making a sturdier form of the virus that will be able to survive once t
  3. 3. and they can bud fine without it so they don’t make it until they need it they only need POLYHEDRIN late in the infection that sturdy shell’s made up of a protein called POLYHEDRIN (and some other stuff) especially since they need a TON of it to make the occlusion bodies early infection very late infection cell hopping host swapping so they have to basically devote all their time & resources to making it and then they make a TON of it PolH gene PolH promoter
  4. 4. we replace the polyH gene with a gene for our protein now the cells start making our protein when they “think” they’re making polyhedrin which they only do in the very late phase but we still need to get it into the cells… so we have an unneeded protein in prime spot for expression…
  5. 5. the bacmid DNA needs the coat to get into cells on its own + transfection reagent some cells take in the bacmid and start making budded virus it has the coat so it can get into the uninfected cells and then those can start making & excreting virus into the liquid they’re in we let the cells make lots of virus bacmid we use cells that are from insects, but we grow them in dishes and later flasks or bags ated cells because it’s harder to get the bacmid in. late so we have to help it out
  6. 6. collect the secreted virus oh no! a cell got in no worries - we’re going to syringe filter it this is the V0. you can keep it in the fridge until you’re ready to use it V0 use it for what? making more virus!
  7. 7. the virus gets into the cells we want to collect that virus. but we need to separate it from the cells. but since the cells aren’t stuck down, we need to centrifuge them to pull them down take that V0 & add it to some cells in a flask now those cells start making virus o a lot of virus. so the next step is to make more. but no V1
  8. 8. there’ll be some protein in here, so you can keep the pellet if you did a biggish V1 but the big yield’s still to come! centrifuge to separate the cells from the virus what we’re after: coated virus V1
  9. 9. use that V1 to infect lots of cells what we’re after: protein-stuffed cells
  10. 10. do the same spin -down but this time focus on the pellet what we’re after: protein-stuffed cells