He jamestheawesome1 thanks for letting me talk about my science to you!
Engineering proteins is way cool, but not quite as crazy as it sounds at first! So, here it is!
– First you get DNA from the organism that you want to make the protein out of. You need to make lots of this DNA so you do a PCR. This uses other proteins (enzymes) to make millions of copies of the DNA. You use a special machine for this calle a thermal cycler. It heats and cools in cycles at very strict timeframes.
– Once you have your DNA from the PCR you make sure it is pure by doing a thing called electrophoresis. This pulls DNA through a solid gel using electricity! You cut the Gel where the DNA is and clean it up using a kit you buy.
– Then you join the DNA into a big circular piece of DNA using enzymes that cut DNA. This is called a plasmid ligation. Once you have your DNA in the plasmid, you can put it into some bacteria. To put it into bacteria, you mix the DNA and bacteria together. Then you either give the bacteria a heat or electric shock. This makes tiny holes open up all over it and it sucks the plasmid inside it! You need a special machine to give it the electric shock.
Now you grow your bacteria! You do this in some special liquid that has all of the essential things a bacteria needs to grow. You keep it in a big fridge that can keep the bacteria at the right temperature (we use 37 and 16 degrees). Once you have grown it you spin it at about 5,000 times per minute in a big centrifuge. Then the fun bit! We break open the bacteria. We do this in a big machine called a French Press. It suqeezes the bacteria at 10,000 psi! The bacteria are broken, and all te proteins that were made inside the bacteria from our plasmid DNA are free!
– Finally, we purify the protein. So we spin the broken bacteria at 40,000 times per min! Then we use special tiny beads made of nickel to bind to our proteins, and we separate the proteins we want to keep (that bind the nuckel) from all of the other proteins in the bacteria.
ALL DONE! You now have an engineered protein you made yourself from DNA. You can also do the same thing to join proteins together by sticking their DNA together in the plasmid! Or cutting bits of proteins up!
I hope I explained it all, sorry it it’s a bit long, but I love my science! If you have any questions about any of the equipment or steps let me know, I can’t wait to answer them!
He jamestheawesome1 thanks for letting me talk about my science to you!
Engineering proteins is way cool, but not quite as crazy as it sounds at first! So, here it is!
– First you get DNA from the organism that you want to make the protein out of. You need to make lots of this DNA so you do a PCR. This uses other proteins (enzymes) to make millions of copies of the DNA. You use a special machine for this calle a thermal cycler. It heats and cools in cycles at very strict timeframes.
– Once you have your DNA from the PCR you make sure it is pure by doing a thing called electrophoresis. This pulls DNA through a solid gel using electricity! You cut the Gel where the DNA is and clean it up using a kit you buy.
– Then you join the DNA into a big circular piece of DNA using enzymes that cut DNA. This is called a plasmid ligation. Once you have your DNA in the plasmid, you can put it into some bacteria. To put it into bacteria, you mix the DNA and bacteria together. Then you either give the bacteria a heat or electric shock. This makes tiny holes open up all over it and it sucks the plasmid inside it! You need a special machine to give it the electric shock.
Now you grow your bacteria! You do this in some special liquid that has all of the essential things a bacteria needs to grow. You keep it in a big fridge that can keep the bacteria at the right temperature (we use 37 and 16 degrees). Once you have grown it you spin it at about 5,000 times per minute in a big centrifuge. Then the fun bit! We break open the bacteria. We do this in a big machine called a French Press. It suqeezes the bacteria at 10,000 psi! The bacteria are broken, and all te proteins that were made inside the bacteria from our plasmid DNA are free!
– Finally, we purify the protein. So we spin the broken bacteria at 40,000 times per min! Then we use special tiny beads made of nickel to bind to our proteins, and we separate the proteins we want to keep (that bind the nuckel) from all of the other proteins in the bacteria.
ALL DONE! You now have an engineered protein you made yourself from DNA. You can also do the same thing to join proteins together by sticking their DNA together in the plasmid! Or cutting bits of proteins up!
I hope I explained it all, sorry it it’s a bit long, but I love my science! If you have any questions about any of the equipment or steps let me know, I can’t wait to answer them!
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