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Team:Lethbridge Canada/project
From 2013hs.igem.org
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<h1>Introduction:</h1> | <h1>Introduction:</h1> | ||
| - | <p>Our project, is to create a natural form of Oxytocin that can be used for widespread research and medical application. Oxytocin has a very short half-life- ranging from five to ten minutes, in its active form. This means that over time, the hormone is subject to degrade very quickly and become unusable. The goal is produce the hormone, attached to it's carrier molecule, neurophysin in order to prevent the breakdown Oxytocin. We hope that our project will be able to make Oxytocin more readily available for study.</p> | + | <p>Our project, is to create a natural form of Oxytocin that can be used for widespread research and medical application. Oxytocin has a very short half-life- ranging from five to ten minutes, in its active form. This means that over time, the hormone is subject to degrade very quickly and become unusable. The goal is produce the hormone, attached to it's carrier molecule, neurophysin in order to prevent the breakdown of Oxytocin. We hope that our project will be able to make Oxytocin more readily available for study.</p> |
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<h1>What is Oxytocin?:</h1> | <h1>What is Oxytocin?:</h1> | ||
| - | <p>Oxytocin is a hormone that has many effects on the body. Physically, it is known to stimulate uterine contractions, aiding the mother in birth. However, Oxytocin also has many other applications with social interaction. Oxytocin helps foster a bond between the mother and child, and stimulates a positive reaction when participating in social interaction. At present, Oxytocin is not comprehensively understood by researchers regarding its effects. Many studies have been undertaken to determine exactly how Oxytocin interacts with the body. In some cases, Oxytocin provides results suggesting that it will enhance the social behaviours of animals and humans when added to their system. It is thought to improve facial recognition between face-to-face interactions, assisting in picking up on emotional cues. If true, Oxytocin could eventually aid people with social bonding disorders, such as autism, schizophrenia, and depression. Yet in some cases, it also produced results indicating individuals would isolate themselves into groups, ignoring any others that are different. Researchers also do not have a strong case towards whether Oxytocin will have positive or negative effects in the human body when used long term, as so far all experiments have only dealt with short term effects on humans. All things considered, having cheap and efficient Oxytocin to study could greatly enhance our knowledge of the hormone, and eventually our ability to treat certain social disorders.</p> | + | <p>Oxytocin is a hormone that has many effects on the body. Physically, it is known to stimulate uterine contractions, aiding the mother in birth. However, Oxytocin also has many other applications with social interaction. Oxytocin helps foster a bond between the mother and child, and stimulates a positive reaction when participating in social interaction. At present, Oxytocin is not comprehensively understood by researchers regarding its wide and varied effects. Many studies have been undertaken to determine exactly how Oxytocin interacts with the body. In some cases, Oxytocin provides results suggesting that it will enhance the social behaviours of animals and humans when added to their system. It is thought to improve facial recognition between face-to-face interactions, assisting in picking up on emotional cues. If true, Oxytocin could eventually aid people with social bonding disorders, such as autism, schizophrenia, and depression. Yet in some cases, it also produced results indicating individuals would isolate themselves into groups, ignoring any others that are different. Researchers also do not have a strong case towards whether Oxytocin will have positive or negative effects in the human body when used long term, as so far all experiments have only dealt with short term effects on humans. All things considered, having cheap and efficient Oxytocin to study could greatly enhance our knowledge of the hormone, and eventually our ability to treat certain social disorders.</p> |
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<h2>Oxytocin-Neurophysin1:</h2> | <h2>Oxytocin-Neurophysin1:</h2> | ||
| - | <p>We accomplish the task of synthesizing | + | <p>We accomplish the task of synthesizing Oxytocin though the use of two separate constructs. The first, is a system designed from maximal gene expression in order to produce the greatest amount of hormone possible. In nature, Oxytocin is naturally produced with its carrier molecule: Neurophysin I. This carrier protein inhibits the degradation of Oxytocin; prolonging its shelf-life. This combined compound is known as prepro-oxyphisin and is the gene of interest expressed in our system.</p> |
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| + | <p>However, some modifications needed to be made to the gene during synthesis. First, a Signal Sequence native to <i>E. coli</i> was added to allow for the protein to be exported beyond the cell due to the lack of Golgi Apparatus within the organism. Additionally, histidine tags were added to the end of the protein to assist with its detection.</p> | ||
<img src="https://static.igem.org/mediawiki/2013hs/8/85/Leth_hs_2013_general_construct_diagram_v2.png" alt="Construct Diagram" /> | <img src="https://static.igem.org/mediawiki/2013hs/8/85/Leth_hs_2013_general_construct_diagram_v2.png" alt="Construct Diagram" /> | ||
Revision as of 16:31, 21 June 2013
Contents:
Introduction:
Our project, is to create a natural form of Oxytocin that can be used for widespread research and medical application. Oxytocin has a very short half-life- ranging from five to ten minutes, in its active form. This means that over time, the hormone is subject to degrade very quickly and become unusable. The goal is produce the hormone, attached to it's carrier molecule, neurophysin in order to prevent the breakdown of Oxytocin. We hope that our project will be able to make Oxytocin more readily available for study.
What is Oxytocin?:
Oxytocin is a hormone that has many effects on the body. Physically, it is known to stimulate uterine contractions, aiding the mother in birth. However, Oxytocin also has many other applications with social interaction. Oxytocin helps foster a bond between the mother and child, and stimulates a positive reaction when participating in social interaction. At present, Oxytocin is not comprehensively understood by researchers regarding its wide and varied effects. Many studies have been undertaken to determine exactly how Oxytocin interacts with the body. In some cases, Oxytocin provides results suggesting that it will enhance the social behaviours of animals and humans when added to their system. It is thought to improve facial recognition between face-to-face interactions, assisting in picking up on emotional cues. If true, Oxytocin could eventually aid people with social bonding disorders, such as autism, schizophrenia, and depression. Yet in some cases, it also produced results indicating individuals would isolate themselves into groups, ignoring any others that are different. Researchers also do not have a strong case towards whether Oxytocin will have positive or negative effects in the human body when used long term, as so far all experiments have only dealt with short term effects on humans. All things considered, having cheap and efficient Oxytocin to study could greatly enhance our knowledge of the hormone, and eventually our ability to treat certain social disorders.
Parts:
Oxytocin-Neurophysin1:
We accomplish the task of synthesizing Oxytocin though the use of two separate constructs. The first, is a system designed from maximal gene expression in order to produce the greatest amount of hormone possible. In nature, Oxytocin is naturally produced with its carrier molecule: Neurophysin I. This carrier protein inhibits the degradation of Oxytocin; prolonging its shelf-life. This combined compound is known as prepro-oxyphisin and is the gene of interest expressed in our system.
However, some modifications needed to be made to the gene during synthesis. First, a Signal Sequence native to E. coli was added to allow for the protein to be exported beyond the cell due to the lack of Golgi Apparatus within the organism. Additionally, histidine tags were added to the end of the protein to assist with its detection.
J23100_B0032_OXT_B0015
NEC1 Enzyme:
As for the cleavage enzyme, Nec I, we constructed multiple promoters in order to find the most efficient way to produce our functional oxytocin when we require it. We will figure this out using our mathematical model where we take into account how many pre-pro oxyphysin will be cut per second per Nec I, how long Nec I lives, etc... Afterwards, we will find the two amounts that correlate best to produce our functional oxytocin without too little or too much cutting enzyme. We have the mathematical model to find out what our projected values are (still in progress) and we have a histidine tag place on our pre-pro oxyphysin so that we can run tests on our constructs.
J231##_B0032_NEC1/PCSK1_B0015
Assays:
Results yet to come! Stay with us.
Math Modeling:
For our project, we attempted to model the protein output of our cells. This would help us in finding the correct ratio of enzyme to protein to express. The full explanation as to how the math model works can be found on our Math Model page.
Conclusion:
In the end, we hope to produce a working Oxytocin-Neurophysin I construct along with a functional NEC I construct. If we manage to attain those two constructs, we can obtain the hormone oxytocin in its natural form. It is our hope that our construct will be able to substantially reduce the cost of producing oxytocin in the commercial environment. Once we have constructed a working system, sending our parts into the parts registry will help future researchers study the effects that oxytocin has on the human system using the naturally occurring form of the hormone.
