And you actually wouldn'tīe able to see it visually but there is E. coli in particular, and so what we could do is, we could, let's say that we have a bunch, let's say you have a vial right over here. And an organism that's typically used, or a type of organism is bacteriaĪnd E. And so now we have this plasmid and we want to insert it into an organism that can make the copies for us. It as helping to do, helping to do the pasting. So that is DNA ligase, which you can think of DNA ligase, to connect the backbones right over here. Right sequences at their ends so that they match up and then you also put inĪ bunch of DNA ligase. They're bumping in just the right way to cause this reaction to happen then you're taking those genes and you're putting them with the plasmids that happen to have the The restriction enzymes are just in mass cutting these things. You're making these solutions and you're applying the And this is amazing because obviously DNA, this isn't stuff that we can, you know, manipulate with our hands the way that we would copyĪnd paste things with tape.
Them to react with each other if they have these overhangs. Pairs over the overhangs, which will allow it easier, it will become easier for And so the plasmid that we're placing in might have complimentary base So you might have an overhang over there, you might have an overhang over there. And in order for them to fit there's oftentimes these And plasmids tend to be circular DNA so we will paste it into a plasmid. So then so this is where we cut, let me write this, we cut out the gene and then we wanna paste it then we wanna paste it into a plasmid. Or it can even express itself just like the genes of the organism that are in the chromosomes,Įxpress themselves. Of genetic material that sits outside of chromosomes but it can reproduce along, or I guess we can say can replicate along with the machinery of the, the genetic machinery of the organism. You've used the restrictionĮnzymes to cut out your gene and then what you wannaĭo is you wanna paste it into what we'll call a plasmid. Left over on either side but essentially you have cut out the gene. And so now you would have, after you applied the restriction enzymes, you will have just that gene. These, those things right over there those are restriction enzymes. Sequence at the other side that we wanna cut. So that might be a restrictionĮnzyme right over there and then you might useĪnother restriction enzyme that identifies with the Sequence right over here and cuts right in the right place. We might use one restriction enzyme, Let me use a different color here, that latches on right over here and identifies the genetic They recognize specific sequences and then we can figure out well which restrictionĮnzyme should we use to cut out different pieces of DNA, but we have gotten to that Identify these enzymes and we know at what points Have gotten to the point that we can find and Restriction enzymes, and I personally find it fascinating that we as a civilization And the way we do that is using restriction enzymes. Well, the first thing we wanna do is we wanna cut this gene out some how. This is the double-stranded DNA and let's say that this part of this DNA has a gene that we want to clone. Actually, let me just draw, let me just try to draw the two strands just so we remind ourselves. I don't want to have to take the trouble of keep drawing the multiple strands. Strand of DNA right over here and I'm just drawing it as a long, but this is a double-stranded, and I'll just write it down, this is double stranded. So how do we do that? Well let's say that this is a But when we talk aboutĬloning and DNA cloning we're talking about somethingĪ little bit simpler. Or an organism, like a sheep, well then you are creating an animal that has the exact genetic Heard the term cloning in terms of the Clone Wars in Star Wars or Dolly the sheep and that is a related idea. And usually it's a piece of DNA that codes for something we care about, it is a gene that willĮxpress itself as a protein that we think is useful in some way. Which is all about making identical copies of a piece of DNA.
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