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So here we have a male that carries, that, has that phenotype or that disease. So that means it's uppercase B and Y. This one must also be X uppercase B and Y. This must be X uppercase B y. Now this individual and this individual can either be this trade or this trade because we have two different possibilities. So let's actually carry out the pudding square to see if these are possible right over here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
This must be X uppercase B y. Now this individual and this individual can either be this trade or this trade because we have two different possibilities. So let's actually carry out the pudding square to see if these are possible right over here. So the choices here are either X uppercase VX uppercase B, or we have X uppercase VX lowercase B. And here the choices are also X uppercase VX uppercase B, or X uppercase VX lowercase B. Okay? | Pedigree Analysis for Autosomal Dominant Traits .txt |
So the choices here are either X uppercase VX uppercase B, or we have X uppercase VX lowercase B. And here the choices are also X uppercase VX uppercase B, or X uppercase VX lowercase B. Okay? So let's move on to this one over here. So we're crossing this one with this one. Let's see what we produce. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So let's move on to this one over here. So we're crossing this one with this one. Let's see what we produce. So let's assume that first, let's assume that the individual is this right over here. If the individual was this, then we have the Xcel. So one type of Xcel, which is that, and then we have this type of sperm cell or this one from the male parent. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So let's assume that first, let's assume that the individual is this right over here. If the individual was this, then we have the Xcel. So one type of Xcel, which is that, and then we have this type of sperm cell or this one from the male parent. And so if we carry out this crossing process, we get X uppercase B, x lowercase B, x uppercase B, x lowercase B. Okay? And actually we don't have to do well, yeah, let's continue, let's finish this up. | Pedigree Analysis for Autosomal Dominant Traits .txt |
And so if we carry out this crossing process, we get X uppercase B, x lowercase B, x uppercase B, x lowercase B. Okay? And actually we don't have to do well, yeah, let's continue, let's finish this up. So X-B-Y-X-B-Y. So from this information, what this tells us is every single individual produced in this case must carry that particular phenotype for that disease. And we know that is not consistent with the information that is given to us because these two individuals don't carry that particular phenotype as described by this pun and square. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So X-B-Y-X-B-Y. So from this information, what this tells us is every single individual produced in this case must carry that particular phenotype for that disease. And we know that is not consistent with the information that is given to us because these two individuals don't carry that particular phenotype as described by this pun and square. So we know it cannot be this genotype here. So, okay, now we know that. Let's continue into the next one. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So we know it cannot be this genotype here. So, okay, now we know that. Let's continue into the next one. Let's suppose the genotype is in fact this one right over here. So now let's carry out the pun and square for that. So we have X uppercase B, we have X lowercase B, and we have X lowercase by for our sperm cells here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
Let's suppose the genotype is in fact this one right over here. So now let's carry out the pun and square for that. So we have X uppercase B, we have X lowercase B, and we have X lowercase by for our sperm cells here. We have X uppercase B. We have x lowercase b, x lowercase b, x lowercase b, x uppercase B-Y-X lowercase b y. Okay, so this actually makes sense because we have a 25 chance that a 25% chance that this individual is formed, which we have right over here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
We have X uppercase B. We have x lowercase b, x lowercase b, x lowercase b, x uppercase B-Y-X lowercase b y. Okay, so this actually makes sense because we have a 25 chance that a 25% chance that this individual is formed, which we have right over here. We have a 25% chance that this individual is formed and a 25% chance that this individual is formed. And that's consistent with these individuals in the following pedigree. So that works as long as this is this individual right over here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
We have a 25% chance that this individual is formed and a 25% chance that this individual is formed. And that's consistent with these individuals in the following pedigree. So that works as long as this is this individual right over here. Okay, now let's move on to this case. We have x uppercase B-Y-X lowercase b. X lowercase b. If we carry out the punning square, we'll see that these individuals do in fact work. | Pedigree Analysis for Autosomal Dominant Traits .txt |
Okay, now let's move on to this case. We have x uppercase B-Y-X lowercase b. X lowercase b. If we carry out the punning square, we'll see that these individuals do in fact work. So because X B can be donated by one parent, XB can be donated by the other parent. So we can in fact form this individual. And likewise, if the Y is donated by the male parent, XB is donated by the other one, we form this individual right over here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So because X B can be donated by one parent, XB can be donated by the other parent. So we can in fact form this individual. And likewise, if the Y is donated by the male parent, XB is donated by the other one, we form this individual right over here. Okay? So to determine if it is sex link dominant, we have to show that all these actually make sense. So we're crossing this individual with this individual. | Pedigree Analysis for Autosomal Dominant Traits .txt |
Okay? So to determine if it is sex link dominant, we have to show that all these actually make sense. So we're crossing this individual with this individual. So what do we get? Well, we have X uppercase B and Y. So from this individual we have X lowercase BX, lowercase B. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So what do we get? Well, we have X uppercase B and Y. So from this individual we have X lowercase BX, lowercase B. And so when we cross, we get X uppercase B, lowercase BX, uppercase BX, lowercase B, and right away we should see that that's a problem. And that's a problem because we form this female right over here. So according to the pedigree, we have this female individual that contains both lowercase B's on both of those X chromosomes. | Pedigree Analysis for Autosomal Dominant Traits .txt |
And so when we cross, we get X uppercase B, lowercase BX, uppercase BX, lowercase B, and right away we should see that that's a problem. And that's a problem because we form this female right over here. So according to the pedigree, we have this female individual that contains both lowercase B's on both of those X chromosomes. But in this particular Punnett square, we see that is impossible because 100% of those females must be heterozygous and so they must express that disease, that phenotype, and that is inconsistent with this particular information. And so it cannot be sex link dominant. So he basically answered our question. | Pedigree Analysis for Autosomal Dominant Traits .txt |
But in this particular Punnett square, we see that is impossible because 100% of those females must be heterozygous and so they must express that disease, that phenotype, and that is inconsistent with this particular information. And so it cannot be sex link dominant. So he basically answered our question. Now we want to show that it could be autosomal dominant. So let's remove all these, all the scrap work that we did to basically show that it wasn't sex linked. And now we're going to follow the same exact step to show that it could be autosomal dominant. | Pedigree Analysis for Autosomal Dominant Traits .txt |
Now we want to show that it could be autosomal dominant. So let's remove all these, all the scrap work that we did to basically show that it wasn't sex linked. And now we're going to follow the same exact step to show that it could be autosomal dominant. So I guess we can hopefully you wrote that information down or you can rewind it if you like. Okay, so we want to now show that it is possible. This pedigree can describe autosomal dominance. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So I guess we can hopefully you wrote that information down or you can rewind it if you like. Okay, so we want to now show that it is possible. This pedigree can describe autosomal dominance. And what that means is, so uppercase B, uppercase B or uppercase B, lower case B will basically describe that disease gene or disease. It will describe, I should say the disease phenotype. Okay? | Pedigree Analysis for Autosomal Dominant Traits .txt |
And what that means is, so uppercase B, uppercase B or uppercase B, lower case B will basically describe that disease gene or disease. It will describe, I should say the disease phenotype. Okay? And the only time we don't have phenotype for the disease is lowercase B, lower case B, okay? So we don't have a disease here. So this must be lowercase B, lower case B, this must be lowercase B, lower case B. | Pedigree Analysis for Autosomal Dominant Traits .txt |
And the only time we don't have phenotype for the disease is lowercase B, lower case B, okay? So we don't have a disease here. So this must be lowercase B, lower case B, this must be lowercase B, lower case B. This must be lowercase b. Lowercase b? So every one that appears normal basically is lowercase B, lowercase B. So this one is lowercase B, and this one is lowercase B as well. | Pedigree Analysis for Autosomal Dominant Traits .txt |
This must be lowercase b. Lowercase b? So every one that appears normal basically is lowercase B, lowercase B. So this one is lowercase B, and this one is lowercase B as well. Now all the ones that have the Z's are either this or that. So this can be either uppercase B, uppercase B, or it can be uppercase B, lowercase B. This can be uppercase B, uppercase B or uppercase B. Lowercase B. | Pedigree Analysis for Autosomal Dominant Traits .txt |
Now all the ones that have the Z's are either this or that. So this can be either uppercase B, uppercase B, or it can be uppercase B, lowercase B. This can be uppercase B, uppercase B or uppercase B. Lowercase B. This one is uppercase B, uppercase B or lowercase B, lower case B. And the same thing with these two. Okay, so these are kind of the possibilities of our genotype. | Pedigree Analysis for Autosomal Dominant Traits .txt |
This one is uppercase B, uppercase B or lowercase B, lower case B. And the same thing with these two. Okay, so these are kind of the possibilities of our genotype. So let's actually pick one and see that they are consistent. So let's begin with these two phenotypes here. So when we cross BB with BB, the only type of phenotype that we form is obviously lowercase B, lowercase B, and that is absolutely consistent with these two individuals here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
So let's actually pick one and see that they are consistent. So let's begin with these two phenotypes here. So when we cross BB with BB, the only type of phenotype that we form is obviously lowercase B, lowercase B, and that is absolutely consistent with these two individuals here. Both of the individuals don't show that disease phenotype and so they're lowercase B, lowercase B. So this actually works out. What about this case? | Pedigree Analysis for Autosomal Dominant Traits .txt |
Both of the individuals don't show that disease phenotype and so they're lowercase B, lowercase B. So this actually works out. What about this case? Well, notice that this individual cannot be uppercase B, uppercase B, because if it were uppercase B uppercase B, what that means is these two individuals would be heterozygous and so that is not true. What about if it's this? Well if it's this, then we have BB, we have lowercase B, lowercase B, and so what this would produce is it would produce 50% heterozygous, so 50% heterozygous and 50% homozygous recessive. | Pedigree Analysis for Autosomal Dominant Traits .txt |
Well, notice that this individual cannot be uppercase B, uppercase B, because if it were uppercase B uppercase B, what that means is these two individuals would be heterozygous and so that is not true. What about if it's this? Well if it's this, then we have BB, we have lowercase B, lowercase B, and so what this would produce is it would produce 50% heterozygous, so 50% heterozygous and 50% homozygous recessive. And that is consistent with our results. We see that the only time this works out is if this individual's upper case B, lowercase B. In that case this works out, this works out and this also works out. | Pedigree Analysis for Autosomal Dominant Traits .txt |
And that is consistent with our results. We see that the only time this works out is if this individual's upper case B, lowercase B. In that case this works out, this works out and this also works out. And this must be this individual, it can't be that homozygous dominant. So now we have uppercase B, lowercase B that is crossed with lowercase B, lowercase B. So let's say uppercase B. Uppercase B. | Pedigree Analysis for Autosomal Dominant Traits .txt |
And this must be this individual, it can't be that homozygous dominant. So now we have uppercase B, lowercase B that is crossed with lowercase B, lowercase B. So let's say uppercase B. Uppercase B. No, that is not true. So now he wants to basically show this one. So we have the male individual, uppercase B, lowercase B, those are the sperm cells and these are the X cells, lowercase B, lowercase B. | Pedigree Analysis for Autosomal Dominant Traits .txt |
No, that is not true. So now he wants to basically show this one. So we have the male individual, uppercase B, lowercase B, those are the sperm cells and these are the X cells, lowercase B, lowercase B. And so we have the following mating process take place and we have heterozygous and we have homozygous recessive. And so these are the possibilities of these offspring right over here. So we have BB, which is consistent with this one and is consistent with this one right over here. | Pedigree Analysis for Autosomal Dominant Traits .txt |
The next big topic that we're going to focus on will be glucose metabolism. And in this lecture, I'd like to basically introduce what this concept actually is, what it involves and what it attempts to actually achieve. So let's begin by describing, generally speaking, what glucose metabolism is. So glucose metabolism is the process by which our body, ourselves, basically try to transform the energy that is stored in the chemical bonds of the sugar molecule, the glucose, into the energy that is stored in the bonds of ATP molecules. Because ultimately it's not the glucose molecules, but the ATP molecules that are used by the cells to carry out many different types of processes. For instance, creating electrochemical gradients by using membrane pumps or using ATP to basically contract muscle. | Introduction to Glucose Metabolism .txt |
So glucose metabolism is the process by which our body, ourselves, basically try to transform the energy that is stored in the chemical bonds of the sugar molecule, the glucose, into the energy that is stored in the bonds of ATP molecules. Because ultimately it's not the glucose molecules, but the ATP molecules that are used by the cells to carry out many different types of processes. For instance, creating electrochemical gradients by using membrane pumps or using ATP to basically contract muscle. So contraction of muscle, it can be skeleton muscle or cardiac muscle or smooth muscle, uses ATP molecules. Now, what types of processes actually make up glucose metabolism? So the first process that we're going to look at will be glycolysis. | Introduction to Glucose Metabolism .txt |
So contraction of muscle, it can be skeleton muscle or cardiac muscle or smooth muscle, uses ATP molecules. Now, what types of processes actually make up glucose metabolism? So the first process that we're going to look at will be glycolysis. And glycolysis is the process by which the glucose molecules present in a cytoplasm are broken down into Pyruvate molecules. So two Pyruvate molecules are net result of two ATP molecules which can be used by the cell to carry out some type of process as well as NADH molecules. And we'll discuss what those are in a future lecture. | Introduction to Glucose Metabolism .txt |
And glycolysis is the process by which the glucose molecules present in a cytoplasm are broken down into Pyruvate molecules. So two Pyruvate molecules are net result of two ATP molecules which can be used by the cell to carry out some type of process as well as NADH molecules. And we'll discuss what those are in a future lecture. But ultimately, glycolysis produces these byproduct molecules we call pyruvates. Now, glycolysis is an anaerobic process and what that means is it does not require oxygen to actually take place. So whether or not we have oxygen doesn't actually matter because glycolysis doesn't actually use oxygen. | Introduction to Glucose Metabolism .txt |
But ultimately, glycolysis produces these byproduct molecules we call pyruvates. Now, glycolysis is an anaerobic process and what that means is it does not require oxygen to actually take place. So whether or not we have oxygen doesn't actually matter because glycolysis doesn't actually use oxygen. Now, in the absence of oxygen under anaerobic conditions, when we don't have plenty of oxygen present inside the cell, these Pyruvate molecules will undergo a process known as fermentation. Now, some cells in nature undergo alcohol fermentation and that produces ethanol from the Pyruvate. Other organisms, for instance, cells of our body undergo lactic acid fermentation and that transforms the Pyruvate into lactic acid or the conjugate base of lactic acid is lactate. | Introduction to Glucose Metabolism .txt |
Now, in the absence of oxygen under anaerobic conditions, when we don't have plenty of oxygen present inside the cell, these Pyruvate molecules will undergo a process known as fermentation. Now, some cells in nature undergo alcohol fermentation and that produces ethanol from the Pyruvate. Other organisms, for instance, cells of our body undergo lactic acid fermentation and that transforms the Pyruvate into lactic acid or the conjugate base of lactic acid is lactate. So inside our body, when the cells of our body don't have enough oxygen, they will take the pyruvates and transform them into lactic acid molecules. Now, what happens if there is oxygen present in the cells of our body? Well, in the case of aerobic conditions, when we have oxygen present in the cell, the Pyruvate molecules will move into the mitochondria of the cell. | Introduction to Glucose Metabolism .txt |
So inside our body, when the cells of our body don't have enough oxygen, they will take the pyruvates and transform them into lactic acid molecules. Now, what happens if there is oxygen present in the cells of our body? Well, in the case of aerobic conditions, when we have oxygen present in the cell, the Pyruvate molecules will move into the mitochondria of the cell. And so this is our mitochondria. And inside the mitochondria, we have processes such as Pyruvate decarboxylation as well as the citric acid cycle and the electron transport chain found on the inner membrane of that mitochondria actually uses those oxygen molecules to ultimately transform the Pyruvate into carbon dioxide molecules as well as ATP molecules. In fact, the majority of ATP molecules produced by our cells are formed in the process that takes place inside the mitochondria. | Introduction to Glucose Metabolism .txt |
And so this is our mitochondria. And inside the mitochondria, we have processes such as Pyruvate decarboxylation as well as the citric acid cycle and the electron transport chain found on the inner membrane of that mitochondria actually uses those oxygen molecules to ultimately transform the Pyruvate into carbon dioxide molecules as well as ATP molecules. In fact, the majority of ATP molecules produced by our cells are formed in the process that takes place inside the mitochondria. Now, this is known as aerobic cellular respiration. An aerobic cellular respiration includes not only the processes that take place in the mitochondria, but also actually includes glycolysis itself. But glycolysis is itself an anaerobic process. | Introduction to Glucose Metabolism .txt |
Now, this is known as aerobic cellular respiration. An aerobic cellular respiration includes not only the processes that take place in the mitochondria, but also actually includes glycolysis itself. But glycolysis is itself an anaerobic process. This takes place regardless of whether or not we actually have o, two molecules present in our cell. Now, let's suppose the cell has plenty of ATP molecules to actually go around and so it doesn't actually want to produce any more ATP molecules. What happens in this situation? | Introduction to Glucose Metabolism .txt |
This takes place regardless of whether or not we actually have o, two molecules present in our cell. Now, let's suppose the cell has plenty of ATP molecules to actually go around and so it doesn't actually want to produce any more ATP molecules. What happens in this situation? So in this situation, because we don't actually want to break down the glucose molecules, we want to store the glucose molecules in a form that basically will not be broken down. And so we take the individual glucose molecules and we transform them into their polysaccharide form we call glycogen. Now, what about these pyruvate molecules and these lactate molecules? | Introduction to Glucose Metabolism .txt |
So in this situation, because we don't actually want to break down the glucose molecules, we want to store the glucose molecules in a form that basically will not be broken down. And so we take the individual glucose molecules and we transform them into their polysaccharide form we call glycogen. Now, what about these pyruvate molecules and these lactate molecules? What happens to them in the case that we have many, many ATP inside our body? So in this case, these pyruvate molecules or lactic acid molecules are transformed back into glucose and then the glucose is stored in a form we call glycogen. And the process by which we transform these pyruvates and lactic acid molecules back into glucose is known as gluconeogenesis. | Introduction to Glucose Metabolism .txt |
What happens to them in the case that we have many, many ATP inside our body? So in this case, these pyruvate molecules or lactic acid molecules are transformed back into glucose and then the glucose is stored in a form we call glycogen. And the process by which we transform these pyruvates and lactic acid molecules back into glucose is known as gluconeogenesis. So glucose means glucose, neo means new molecules, and genesis means the formation. So the formation of these new glucose molecules by using the pyruvate or the lactic acid molecules. Now we see that glycolysis breaks down the glucose, but gluconeogenesis uses these byproducts to reform that glucose molecule. | Introduction to Glucose Metabolism .txt |
So glucose means glucose, neo means new molecules, and genesis means the formation. So the formation of these new glucose molecules by using the pyruvate or the lactic acid molecules. Now we see that glycolysis breaks down the glucose, but gluconeogenesis uses these byproducts to reform that glucose molecule. So one goes this way and the other one goes in reverse. So if the cell needs to break down glucose and produce ATP molecules, then what it does is basically shuts off gluconeogenesis. But if the cell has plenty amounts of ATP molecules and it doesn't actually want to break down the glucose, then gluconeogenesis is more likely to actually take place. | Introduction to Glucose Metabolism .txt |
So one goes this way and the other one goes in reverse. So if the cell needs to break down glucose and produce ATP molecules, then what it does is basically shuts off gluconeogenesis. But if the cell has plenty amounts of ATP molecules and it doesn't actually want to break down the glucose, then gluconeogenesis is more likely to actually take place. In fact, we see that glycolysis and gluconeogenesis do not actually take place at the same exact moment in time. So when one process is activated, the other process is usually inhibited and vice versa. So now we know the general idea of what glucose metabolism actually is. | Introduction to Glucose Metabolism .txt |
In fact, we see that glycolysis and gluconeogenesis do not actually take place at the same exact moment in time. So when one process is activated, the other process is usually inhibited and vice versa. So now we know the general idea of what glucose metabolism actually is. But how does that glucose actually make its way into the cells of our body? Or more generally, how does the glucose actually make its way into our body in the first place? Well, via the ingestion of food. | Introduction to Glucose Metabolism .txt |
But how does that glucose actually make its way into the cells of our body? Or more generally, how does the glucose actually make its way into our body in the first place? Well, via the ingestion of food. So if we eat a meal that is rich in carbohydrates, that's how the glucose actually makes its way into our body. So there are two types of sugar molecules carbohydrates that we typically ingest. So carbohydrates polysaccharides that come from plants, and carbohydrates polysaccharides that come from animals. | Introduction to Glucose Metabolism .txt |
So if we eat a meal that is rich in carbohydrates, that's how the glucose actually makes its way into our body. So there are two types of sugar molecules carbohydrates that we typically ingest. So carbohydrates polysaccharides that come from plants, and carbohydrates polysaccharides that come from animals. Now, for instance, if we eat a piece of chicken, that chicken not only has protein and fat, it also contains polysaccharides carbohydrates store in a form we call glycogen, which is the same form that we mentioned just a moment ago. Now, if we ingest things like pasta or bread or cereal, these are actually polysaccharides that come from plants. And so what we're ingesting is starch. | Introduction to Glucose Metabolism .txt |
Now, for instance, if we eat a piece of chicken, that chicken not only has protein and fat, it also contains polysaccharides carbohydrates store in a form we call glycogen, which is the same form that we mentioned just a moment ago. Now, if we ingest things like pasta or bread or cereal, these are actually polysaccharides that come from plants. And so what we're ingesting is starch. And there are two types of starch. So we have amylose and amylopectin. So one of them is basically a linear helical structure that's the amylose. | Introduction to Glucose Metabolism .txt |
And there are two types of starch. So we have amylose and amylopectin. So one of them is basically a linear helical structure that's the amylose. And the amylopectin is like the glycogen, actually a branched form of starch. Now, we see that these polysaccharides are inherently too large to actually fit into our cells, and they're too large to actually move around and transport in the blood plasma. And so before these large polysaccharides actually make their way into the blood plasma of our body and into our cells, these carbohydrates must be broken down into smaller components. | Introduction to Glucose Metabolism .txt |
And the amylopectin is like the glycogen, actually a branched form of starch. Now, we see that these polysaccharides are inherently too large to actually fit into our cells, and they're too large to actually move around and transport in the blood plasma. And so before these large polysaccharides actually make their way into the blood plasma of our body and into our cells, these carbohydrates must be broken down into smaller components. In fact, they must be broken down into these individual glucose molecules before the cells can actually uptake those glucose molecules and store the glucose as glycogen, or break down the glucose to form ATP molecules. So the question is, what are these enzymes, digestive enzymes, proteases, that basically break down these carbohydrates into their individual monomeric form? Well, we have many different types of enzymes, and I've listed six enzymes, actually seven enzymes on the board. | Introduction to Glucose Metabolism .txt |
In fact, they must be broken down into these individual glucose molecules before the cells can actually uptake those glucose molecules and store the glucose as glycogen, or break down the glucose to form ATP molecules. So the question is, what are these enzymes, digestive enzymes, proteases, that basically break down these carbohydrates into their individual monomeric form? Well, we have many different types of enzymes, and I've listed six enzymes, actually seven enzymes on the board. And let's begin with a salivary alpha amylase. So salivary simply means it exists in a saliva. So when we eat food and where we're chewing the food, that saliva actually contains a specific type of digestive enzyme and proteins known as alpha amylase. | Introduction to Glucose Metabolism .txt |
And let's begin with a salivary alpha amylase. So salivary simply means it exists in a saliva. So when we eat food and where we're chewing the food, that saliva actually contains a specific type of digestive enzyme and proteins known as alpha amylase. And what the alpha amylase does is it begins to cleave alpha one four glycocity linkages that exist in the starch as well as glycogen. So this begins the cleavage of alpha one four glycocity linkages in the mouth, and this breaks it down to smaller polysaccharides and oligosaccharides. Now, from the mouth, it moves via the esophagus, eventually makes its way into our stomach. | Introduction to Glucose Metabolism .txt |
And what the alpha amylase does is it begins to cleave alpha one four glycocity linkages that exist in the starch as well as glycogen. So this begins the cleavage of alpha one four glycocity linkages in the mouth, and this breaks it down to smaller polysaccharides and oligosaccharides. Now, from the mouth, it moves via the esophagus, eventually makes its way into our stomach. Now, in the stomach, nothing actually breaks down. So what that means is the actual glycocitic linkages, the bonds don't break down in the stomach, but once it makes its way into the small intestine, that's when the rest of that digestion actually takes place, because the pancreas produces a specific type of carbohydrate digestive enzyme known as pancreatic alpha amylase. And this is much more potent and much more powerful than the salivary alpha amylase. | Introduction to Glucose Metabolism .txt |
Now, in the stomach, nothing actually breaks down. So what that means is the actual glycocitic linkages, the bonds don't break down in the stomach, but once it makes its way into the small intestine, that's when the rest of that digestion actually takes place, because the pancreas produces a specific type of carbohydrate digestive enzyme known as pancreatic alpha amylase. And this is much more potent and much more powerful than the salivary alpha amylase. So the pancreatic alpha amylase also is able to break down those same alpha one four glycocytic linkages, but this actually breaks down the polysaccharide into either disaccharides or trisaccharides. So in the case of starch or glycogen, we basically break down these individual polysaccharides into maltose molecules, which are disaccharides. That consists of two glucose or trisaccharides, that consists of three glucose known as maltotrios. | Introduction to Glucose Metabolism .txt |
So the pancreatic alpha amylase also is able to break down those same alpha one four glycocytic linkages, but this actually breaks down the polysaccharide into either disaccharides or trisaccharides. So in the case of starch or glycogen, we basically break down these individual polysaccharides into maltose molecules, which are disaccharides. That consists of two glucose or trisaccharides, that consists of three glucose known as maltotrios. Now, the actual cells on the epithelium of the small intestine actually contain these secretory vesicles, these granules that themselves contain enzymes. And so these are the enzymes that they basically have. So we have Maltase, we have alpha glucosedase, we have alpha dextrinase, we have sucrase and lactase. | Introduction to Glucose Metabolism .txt |
Now, the actual cells on the epithelium of the small intestine actually contain these secretory vesicles, these granules that themselves contain enzymes. And so these are the enzymes that they basically have. So we have Maltase, we have alpha glucosedase, we have alpha dextrinase, we have sucrase and lactase. And all of these enzymes are basically specific to the type of molecules and type of bonds they actually cleave. So, for instance, in the case of Maltase, maltase is released by the cells on the brush border, and this enzyme basically breaks down the Maltose. So here we said the pancreatic alpha amylase breaks down the ligosaccharides and polysaccharides that could not be broken down by the salivary alpha amylase into Maltose or Maltotria. | Introduction to Glucose Metabolism .txt |
And all of these enzymes are basically specific to the type of molecules and type of bonds they actually cleave. So, for instance, in the case of Maltase, maltase is released by the cells on the brush border, and this enzyme basically breaks down the Maltose. So here we said the pancreatic alpha amylase breaks down the ligosaccharides and polysaccharides that could not be broken down by the salivary alpha amylase into Maltose or Maltotria. And these Maltose molecules are broken down by these maltase enzymes at the brush border of our epithelium of the small intestine. And once the Maltose is broken down into the glucose constituents, then the glucose can actually be taken by the cell by using a special type of glucose transporter, as we'll discuss in a future lecture. Now, we also mentioned the meltotrios, and we have another type of enzyme, a different enzyme known as alpha glucose, a dase, that basically breaks down the meltotrials into the three constituent glucose molecules. | Introduction to Glucose Metabolism .txt |
And these Maltose molecules are broken down by these maltase enzymes at the brush border of our epithelium of the small intestine. And once the Maltose is broken down into the glucose constituents, then the glucose can actually be taken by the cell by using a special type of glucose transporter, as we'll discuss in a future lecture. Now, we also mentioned the meltotrios, and we have another type of enzyme, a different enzyme known as alpha glucose, a dase, that basically breaks down the meltotrials into the three constituent glucose molecules. And only then can the glucose molecules can actually make their way into the cell of our body. Then we also have alpha dextronose or dextrinase. Now, in the case of alpha dextronate, so let's go back to starch and glycogen. | Introduction to Glucose Metabolism .txt |
And only then can the glucose molecules can actually make their way into the cell of our body. Then we also have alpha dextronose or dextrinase. Now, in the case of alpha dextronate, so let's go back to starch and glycogen. So if we discuss so if we ingest the amylopectin version of starch or glycogen, we know that these two types of polysaccharides not only have the alpha one four glycocity bonds, they also have the alpha one six glycocytic bonds. And the alpha amylase found in our mouth and the alpha amylase found in our small intestine that is produced by the pancreas, these cannot break down those alpha one six linkages. And that's where this alpha dextronase actually comes into play. | Introduction to Glucose Metabolism .txt |
So if we discuss so if we ingest the amylopectin version of starch or glycogen, we know that these two types of polysaccharides not only have the alpha one four glycocity bonds, they also have the alpha one six glycocytic bonds. And the alpha amylase found in our mouth and the alpha amylase found in our small intestine that is produced by the pancreas, these cannot break down those alpha one six linkages. And that's where this alpha dextronase actually comes into play. So the alpha dextronase basically breaks down the limited xray, which is basically those oligosaccharides that contain the alpha one six bonds which were not broken down by either of these two types of enzymes. And so it's this one that breaks down these dextrin molecules, breaks down those alpha one six linkages, breaks the molecules into their individual constituent glucose molecules, and then the glucose is ingested into our cell. Now, glucose molecules are not the only sugar molecules that we actually ingest into our body. | Introduction to Glucose Metabolism .txt |
So the alpha dextronase basically breaks down the limited xray, which is basically those oligosaccharides that contain the alpha one six bonds which were not broken down by either of these two types of enzymes. And so it's this one that breaks down these dextrin molecules, breaks down those alpha one six linkages, breaks the molecules into their individual constituent glucose molecules, and then the glucose is ingested into our cell. Now, glucose molecules are not the only sugar molecules that we actually ingest into our body. We can also ingest, for instance, galactase we can ingest or galactose we can ingest fructose and so forth. And so we have many other examples of enzymes that are used to break down these specific types of glycosytic bonds. So we have Succeed, which basically breaks down the glycocitic bond between fructose and sugar and fructose and glucose. | Introduction to Glucose Metabolism .txt |
We can also ingest, for instance, galactase we can ingest or galactose we can ingest fructose and so forth. And so we have many other examples of enzymes that are used to break down these specific types of glycosytic bonds. So we have Succeed, which basically breaks down the glycocitic bond between fructose and sugar and fructose and glucose. So when fructose and glucose combined, they form sucrose, and sucrase breaks down sucrose. So sucrose is essentially a mobile form of a carbohydrate sugar molecule found inside plants. So when we eat plants, we can also actually eat these sucrose molecules. | Introduction to Glucose Metabolism .txt |
So when fructose and glucose combined, they form sucrose, and sucrase breaks down sucrose. So sucrose is essentially a mobile form of a carbohydrate sugar molecule found inside plants. So when we eat plants, we can also actually eat these sucrose molecules. And so Succeed is responsible for breaking down sucrose. Now, we also have lactase. So lactase is essentially a digestive enzyme that breaks down lactose. | Introduction to Glucose Metabolism .txt |
And so Succeed is responsible for breaking down sucrose. Now, we also have lactase. So lactase is essentially a digestive enzyme that breaks down lactose. And lactose is a disaccharide that consists of glucose and galactose. And lactose we obtain from dairy products, from milk. So if we drink milk inside milk, we'll find these lactose disaccharides. | Introduction to Glucose Metabolism .txt |
And lactose is a disaccharide that consists of glucose and galactose. And lactose we obtain from dairy products, from milk. So if we drink milk inside milk, we'll find these lactose disaccharides. And it's the lactose that breaks down these disaccharides into their individual monomers. So once all these different types of enzymes and many more, they basically break down all the different types of carbohydrates inside the small intestine. Only then can these actually make their way into the cytoplasm of our cells and into the blood via this process of transport. | Introduction to Glucose Metabolism .txt |
Now, what's the process by which we can actually sequence our DNA molecule? Well, the process is known as Sanger dioxine method, or simply Sanger DNA sequencing. Now, before we discuss the steps of this process, let's discuss an important molecule used in this process and let's see why it is actually used. So the molecule is this molecule here. It's called two prime, three prime dioxy nucleuside triphosphate or simply Ddntp. Now, this molecule is almost identical to a normal deoxy nucleuside triphosphate. | Sanger Sequencing of DNA .txt |
So the molecule is this molecule here. It's called two prime, three prime dioxy nucleuside triphosphate or simply Ddntp. Now, this molecule is almost identical to a normal deoxy nucleuside triphosphate. The only difference is the sugar component contains a three prime carbon that does not contain a hydroxyl group. So remember, in a normal deoxy nucleus triphosphate, the presence of the hydroxyl group on the three prime carbon allows DNA polymerase to actually form a phosphodiastor bond with the next nucleotide in line. So in the process of DNA synthesis, when we're replicating a DNA strand, the DNA polymerase needs this hydroxyl group to be present on the three prime carbon to actually form the phosphodiaester bond. | Sanger Sequencing of DNA .txt |
The only difference is the sugar component contains a three prime carbon that does not contain a hydroxyl group. So remember, in a normal deoxy nucleus triphosphate, the presence of the hydroxyl group on the three prime carbon allows DNA polymerase to actually form a phosphodiastor bond with the next nucleotide in line. So in the process of DNA synthesis, when we're replicating a DNA strand, the DNA polymerase needs this hydroxyl group to be present on the three prime carbon to actually form the phosphodiaester bond. And if that hydroxyl group is not present, as in this case, it will not be able to form that phospholdiaester bond. And so DNA replication would essentially stop. And so what this DD NTP molecule is used for in this method is to basically stop the process of DNA replication. | Sanger Sequencing of DNA .txt |
And if that hydroxyl group is not present, as in this case, it will not be able to form that phospholdiaester bond. And so DNA replication would essentially stop. And so what this DD NTP molecule is used for in this method is to basically stop the process of DNA replication. And we'll see why that's important towards the end of this lecture. So let's move on to these four steps. So, in step one of the standard DNA sequencing, we have to actually obtain that DNA molecule that we want to sequence. | Sanger Sequencing of DNA .txt |
And we'll see why that's important towards the end of this lecture. So let's move on to these four steps. So, in step one of the standard DNA sequencing, we have to actually obtain that DNA molecule that we want to sequence. So let's suppose we have a double stranded DNA molecule as shown on the board. Now, the second step of this process will involve DNA replication. And remember, DNA replication only takes place if the two strands of DNA have separated. | Sanger Sequencing of DNA .txt |
So let's suppose we have a double stranded DNA molecule as shown on the board. Now, the second step of this process will involve DNA replication. And remember, DNA replication only takes place if the two strands of DNA have separated. So, in step one, what we essentially want to do is we want to denature the double helix structure of the DNA. We want to separate the two strands of DNA, and the way that we're going to separate them is by adding sodium hydroxide. So remember, a base or an acid, if we mix the DNA in either a basic or acidic solution, in this case a basic, the base will essentially ionize the bases of our DNA molecule and that will disrupt and break the hydrogen bonds. | Sanger Sequencing of DNA .txt |
So, in step one, what we essentially want to do is we want to denature the double helix structure of the DNA. We want to separate the two strands of DNA, and the way that we're going to separate them is by adding sodium hydroxide. So remember, a base or an acid, if we mix the DNA in either a basic or acidic solution, in this case a basic, the base will essentially ionize the bases of our DNA molecule and that will disrupt and break the hydrogen bonds. And so if we take the double strand DNA molecule and we add sodium hydroxide, we produce these two individual strands of DNA. Now, one of these single strands of DNA can actually be chosen for the sequencing process. Now, it doesn't matter which one of these DNA strands we choose, because if we choose this one, for example, then once we determine the sequence of this DNA strand, we can easily determine what the sequence of the other strand is because these two strands are complementary. | Sanger Sequencing of DNA .txt |
And so if we take the double strand DNA molecule and we add sodium hydroxide, we produce these two individual strands of DNA. Now, one of these single strands of DNA can actually be chosen for the sequencing process. Now, it doesn't matter which one of these DNA strands we choose, because if we choose this one, for example, then once we determine the sequence of this DNA strand, we can easily determine what the sequence of the other strand is because these two strands are complementary. They have complementary base pairing. So the G bases with our C and the A bases with our T. So once we know this sequence, we know what the other sequence is simply by the base parent process. So let's choose this single stranded DNA molecule. | Sanger Sequencing of DNA .txt |
They have complementary base pairing. So the G bases with our C and the A bases with our T. So once we know this sequence, we know what the other sequence is simply by the base parent process. So let's choose this single stranded DNA molecule. We isolate it, we place it into our beaker that contains only this molecule here. And then we move on to step number two. And in step number two, we want to basically replicate this DNA molecule. | Sanger Sequencing of DNA .txt |
We isolate it, we place it into our beaker that contains only this molecule here. And then we move on to step number two. And in step number two, we want to basically replicate this DNA molecule. And so what that means is we need three different things. We need a DNA primer, we need DNA polymerase and we need the building blocks, the four types of normal deoxy nucleotide triphosphate. So in step two, the solution containing the single strand of DNA, this one here is mixed with number one or a a labeled radioactively labeled DNA primer. | Sanger Sequencing of DNA .txt |
And so what that means is we need three different things. We need a DNA primer, we need DNA polymerase and we need the building blocks, the four types of normal deoxy nucleotide triphosphate. So in step two, the solution containing the single strand of DNA, this one here is mixed with number one or a a labeled radioactively labeled DNA primer. So we need the DNA primer basically for that DNA polymerase to actually work. Because remember, the DNA polymerase will only initiate replication if the primer is present. And what that means is we have to know what this initial sequence is on that DNA molecule. | Sanger Sequencing of DNA .txt |
So we need the DNA primer basically for that DNA polymerase to actually work. Because remember, the DNA polymerase will only initiate replication if the primer is present. And what that means is we have to know what this initial sequence is on that DNA molecule. Because to build the DNA primer, we have to know what the complementary sequence is to this group here. So if this is ACG, then we have to build a primer that contains a sequence TGC. And so we can build that in a laboratory. | Sanger Sequencing of DNA .txt |
Because to build the DNA primer, we have to know what the complementary sequence is to this group here. So if this is ACG, then we have to build a primer that contains a sequence TGC. And so we can build that in a laboratory. And we also radioactively label that DNA primer because that will basically allow us to pinpoint exactly where that molecule is when we undergo the process of gel electrophoresis. So we add a label DNA primer that is complementary to the three end of that single stranded DNA molecule that we want to sequence. So this is the three end of that DNA molecule. | Sanger Sequencing of DNA .txt |
And we also radioactively label that DNA primer because that will basically allow us to pinpoint exactly where that molecule is when we undergo the process of gel electrophoresis. So we add a label DNA primer that is complementary to the three end of that single stranded DNA molecule that we want to sequence. So this is the three end of that DNA molecule. And remember, DNA polymerase reads from three to five and it builds from five to three. And that's why this is the end that we actually want to build the DNA primer for. So we add the DNA polymerase and we also add the four types of deoxy nucleus diet triphosphate. | Sanger Sequencing of DNA .txt |
And remember, DNA polymerase reads from three to five and it builds from five to three. And that's why this is the end that we actually want to build the DNA primer for. So we add the DNA polymerase and we also add the four types of deoxy nucleus diet triphosphate. So we add dATP dGTP dCTP and TTP. And finally, the important component in the standard died oxy method is a tiny amount of one of the four types of Ddntp molecules. Remember, we have four different types that can exist and that's because we have four different types of bases. | Sanger Sequencing of DNA .txt |
So we add dATP dGTP dCTP and TTP. And finally, the important component in the standard died oxy method is a tiny amount of one of the four types of Ddntp molecules. Remember, we have four different types that can exist and that's because we have four different types of bases. So this base could be Adamine, it could be Guanine, it could be Cytosine, or it could be Thymine. And so we have four different types of Ggntp molecules. And in step two, we have to add a tiny amount, about 1% with respect to the other nucleoside triphosphates of a specific dig deoxy nucleuside triphosphate. | Sanger Sequencing of DNA .txt |
So this base could be Adamine, it could be Guanine, it could be Cytosine, or it could be Thymine. And so we have four different types of Ggntp molecules. And in step two, we have to add a tiny amount, about 1% with respect to the other nucleoside triphosphates of a specific dig deoxy nucleuside triphosphate. So we don't add all four types, we only add one type. Now, why is that important? Well, let's see what that actually does by examining the following diagram. | Sanger Sequencing of DNA .txt |
So we don't add all four types, we only add one type. Now, why is that important? Well, let's see what that actually does by examining the following diagram. So we essentially take this DNA molecule, the single strand, and we mix it with these four components. So we have the radioactively labeled DNA primer that is complementary to the three end. We have the four types of deoxy nucleuside triphosphates. | Sanger Sequencing of DNA .txt |
So we essentially take this DNA molecule, the single strand, and we mix it with these four components. So we have the radioactively labeled DNA primer that is complementary to the three end. We have the four types of deoxy nucleuside triphosphates. These four ones shown here, we have the DNA polymerase, and we have a very small amount. So let's say about 1% of the Ddatp. So that's the specific Ddntp that we're going to choose for this specific experiment, for that specific beaker. | Sanger Sequencing of DNA .txt |
These four ones shown here, we have the DNA polymerase, and we have a very small amount. So let's say about 1% of the Ddatp. So that's the specific Ddntp that we're going to choose for this specific experiment, for that specific beaker. Now, what will begin to happen? Well, what will happen is the complementary DNA primer will hybridize itself to this section here as shown in the following diagram. So this is our DNA primer. | Sanger Sequencing of DNA .txt |
Now, what will begin to happen? Well, what will happen is the complementary DNA primer will hybridize itself to this section here as shown in the following diagram. So this is our DNA primer. It will form these base pairs as shown in the following diagram. So T base pairs with a G, base pairs with C, and C base pairs with G. And then the DNA polymerase will bind onto the primer and it will use the hydroxyl group on this cytosine to basically form that first phosphodiaester bond. And so it will take a thymine. | Sanger Sequencing of DNA .txt |
It will form these base pairs as shown in the following diagram. So T base pairs with a G, base pairs with C, and C base pairs with G. And then the DNA polymerase will bind onto the primer and it will use the hydroxyl group on this cytosine to basically form that first phosphodiaester bond. And so it will take a thymine. It will take this molecule here to basically form the first base. Then it will move on onto the second base, which is a T, and that will basically form an A. Now, we have two types of A's that we can use. | Sanger Sequencing of DNA .txt |
It will take this molecule here to basically form the first base. Then it will move on onto the second base, which is a T, and that will basically form an A. Now, we have two types of A's that we can use. One of them is the normal deoxy adenosine triphosphate. The other one is a dioxy adenosine triphosphate. And the dioxia adenosine triphosphate does not contain a hydroxyl group on the third prime carbon. | Sanger Sequencing of DNA .txt |
One of them is the normal deoxy adenosine triphosphate. The other one is a dioxy adenosine triphosphate. And the dioxia adenosine triphosphate does not contain a hydroxyl group on the third prime carbon. And what that means is if that DNA polymerase actually uses the Ddatp to place this base, it will not be able to continue that DNA replication process because that molecule does not have the hydroxyl group that is needed to produce the phosphor diester bond with the next base. And so if this A comes from the Ddatp, this process will end and this will be the molecule that we will synthesize. And this is why we have fragment number one, molecule number one. | Sanger Sequencing of DNA .txt |
And what that means is if that DNA polymerase actually uses the Ddatp to place this base, it will not be able to continue that DNA replication process because that molecule does not have the hydroxyl group that is needed to produce the phosphor diester bond with the next base. And so if this A comes from the Ddatp, this process will end and this will be the molecule that we will synthesize. And this is why we have fragment number one, molecule number one. Now, because we only have 1% of this Ddatp, this DNA polymerase will only sometimes use the Ddatp. Usually it's going to use the normal dATP molecule. And if it uses the normal dATP that contains the hydroxyl, then it will continue synthesizing those fossil diester bonds and so we can produce fragment number two. | Sanger Sequencing of DNA .txt |
Now, because we only have 1% of this Ddatp, this DNA polymerase will only sometimes use the Ddatp. Usually it's going to use the normal dATP molecule. And if it uses the normal dATP that contains the hydroxyl, then it will continue synthesizing those fossil diester bonds and so we can produce fragment number two. So if this was normal, then it will continue. So the DNA polymerase would add the thymine, then the guanine, then the cytosine, then the thiamine. And now it comes to a T. So that means there is once again the possibility for an A. | Sanger Sequencing of DNA .txt |
So if this was normal, then it will continue. So the DNA polymerase would add the thymine, then the guanine, then the cytosine, then the thiamine. And now it comes to a T. So that means there is once again the possibility for an A. And the A can either come from this normal dATP or the abnormal Ddatp that lacks the hydroxyl. And if it's this group here, then once again we will stop the synthesis and this fragment will be produced. Now, if it wasn't that molecule, then we would add the next. | Sanger Sequencing of DNA .txt |
And the A can either come from this normal dATP or the abnormal Ddatp that lacks the hydroxyl. And if it's this group here, then once again we will stop the synthesis and this fragment will be produced. Now, if it wasn't that molecule, then we would add the next. So if the A was normal, the normal dATP, it would produce the next nucleotide in line. And so the next one is also an A because this is a T. So once again, this is an A. And now we have a possibility between this one or this one. | Sanger Sequencing of DNA .txt |
So if the A was normal, the normal dATP, it would produce the next nucleotide in line. And so the next one is also an A because this is a T. So once again, this is an A. And now we have a possibility between this one or this one. And let's suppose it's once again the ggatp. And so it will stop it once again after this because it lacks that hydroxyl. And so at the end in our mixture, in beaker number one, after we conduct step number two with the DD ATP, these are the other three fragments that will be present inside our beaker number one. | Sanger Sequencing of DNA .txt |
And let's suppose it's once again the ggatp. And so it will stop it once again after this because it lacks that hydroxyl. And so at the end in our mixture, in beaker number one, after we conduct step number two with the DD ATP, these are the other three fragments that will be present inside our beaker number one. Now we take that beaker number one and we place it into SDS page. So SDS polyacrylamide gel electrophoresis. So this is our setup, and we have four different wells. | Sanger Sequencing of DNA .txt |
Now we take that beaker number one and we place it into SDS page. So SDS polyacrylamide gel electrophoresis. So this is our setup, and we have four different wells. Now, well, number one, we label as the Ddatp, because this is step two, where we use the DD ATP. We take the mixture and we place it into well number one, lane number one. And we allow these to separate based on their masses. | Sanger Sequencing of DNA .txt |
Now, well, number one, we label as the Ddatp, because this is step two, where we use the DD ATP. We take the mixture and we place it into well number one, lane number one. And we allow these to separate based on their masses. Remember, in gel electrophoresis, the smaller our molecule is, the farther down it will move along that gel. So if this is molecule one, Two and Three, this band will be for molecule One. This band will be for molecule Two. | Sanger Sequencing of DNA .txt |
Remember, in gel electrophoresis, the smaller our molecule is, the farther down it will move along that gel. So if this is molecule one, Two and Three, this band will be for molecule One. This band will be for molecule Two. And this band, the highest up, will be the largest molecule, molecule Three. Now, we continue the same process three more times. And the second time around, we use a different Dgntp. | Sanger Sequencing of DNA .txt |
And this band, the highest up, will be the largest molecule, molecule Three. Now, we continue the same process three more times. And the second time around, we use a different Dgntp. The third time around, we use yet another ggntp. And the final fourth time, we use the final Dgntp. So let's suppose the second time around, instead of using the DG ATP, we used Ddgtp. | Sanger Sequencing of DNA .txt |
The third time around, we use yet another ggntp. And the final fourth time, we use the final Dgntp. So let's suppose the second time around, instead of using the DG ATP, we used Ddgtp. And so instead of having the fragments where we stopped on the A's, we're going to have the fragments where we stop on the g's. And so, because we only have two CS, so this C doesn't count because it's part of the primer. So we have one C and we have a second C. So the complementary would have a g here and a g here. | Sanger Sequencing of DNA .txt |