Contents:

• Introduction - Why this project?
• What is Oxytocin?
• Parts
  • Oxytocin-Neurophysin 1 Construct/Gene
  • Nec 1 Construct/Gene
• Assays/Promoters - Statistical Results
• Math Modeling
• Conclusion

Introduction:

Our project, is to create a natural form of Oxytocin that can be used for widespread research and medical application. All Oxytocin has a very short half-life- ranging from five to ten minutes, in its active form. The goal is produce the hormone, attached to it's carrier molecule, neurophysin. 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 contractions during the birth of children and aids 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. 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, picking up on emotional cues. The effects of Oxytocin could eventually aid people with social bonding disorders, such as autism and depression. Yet in some cases, it also produced negative results causing individuals to 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 all experiments so far only deal with short term effects on humans. With all of this, 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 natural oxytocin by creating two separate constructs. Because oxytocin by itself only has a half-life of <10mins a carrier protein is needed in-order to stabilize our hormone for storage. We accomplish this by using Neurophysin I (a carrier molecule) bound to our Oxytocin molecule to create pre-pro oxyphysin, which has a far longer half-life than oxytocin. Once the Oxytocin-Neurophysin I construct is created - effectively stabilizing our hormone – our E. Coli cells can be stored for when production of the hormone is needed. To separate oxytocin from Neurophysin I we use Nec I (aka PCSK1) which cuts in between our oxytocin and Neurophysin molecules to release oxytocin in its functional form.

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.