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Team:WarrenCentral WCC IN
From 2012hs.igem.org
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Team
Tell us about your team, your school!
-Diamond Jones
Diamond is planning on majoring in genetics at Purdue University. She is actively involved in her school’s honors band and National Honors Society. Diamond has put in countless hours of volunteer work at the Indianapolis Humane Society, churches, retirement homes, and local food pantries. Her future plans include becoming a genetic counselor, and creating a gene that allows her to live for eternity.
-Sarah Perun
Sarah will be studying biology and following the pre-med track at Purdue University. She enjoys traveling, eating, sports, music, reading, and shopping. Sarah is involved with the varsity golf team, band, and National Honors Society. Boiler up!
-Myka Norris
Myka is planning on majoring in nursing at IUPUI. Most of her time is taken up by her job at Steak n Shake. However, in her free time she enjoys running and hanging out with family and friends.
-Rebekah Hodge
Bekah is currently a captain of the Warrren Central Speech team and member of Student Council, Latin club, National Junior Classical League, and Indiana Junior Classical League. She also works part time as a waitress for MCL Restaurant & Bakery. She is planning on attending Indiana University – Bloomington and majoring in Biology/PreMed. Eventually, she hopes to become a pediatrician and run her own family practice.
-Patrick Steuer
The year was 1994, it was a warm spring evening in the month of April, the 11th if I recall correctly, when Patrick Steuer came screaming into the world, changing it in a way it would never truly recover from. Patrick doesn’t do a whole lot aside from chilling, but he does it extremely well. He is going to IU and plans on majoring in human biology. His goal is to eventually become a genetic counselor.
-Lewis Woods
Lewis Manning Woods IV. is a senior and plans on attending IU Bloomington in the fall. He has not yet declared his major but wants to go to medical school, be a doctor and have his own practice when he gets out of school. Outside of school he is very active in his church, and captained the High School soccer team. He also plays tennis for recreation.
-Cheyenne Harvey
Cheyenne is a senior and a dedicated member of Student Council. When she isn't doing volunteer work, she spends most of her time taking care of her beautiful puppy, Tink. Cheyenne plans on attending the University of Indianapolis in the fall where she will be majoring in nursing. In the event that her career as a professional rapper falls through, Cheyenne hopes to instead become an Intensive Care Unit nurse practitioner.
-Brandon Whatley
Brandon Whatley is a top-notch student at Warren Central. He is an active member in student council, the reigning president of the Spanish Club, a member of Connection Show Choir, a member of the Fun Monkey Improv team, and a golf player. He plans on attending Indiana University in Bloomington in the Fall of 2012 and majoring in biology. He also enjoys long walks on the beach and the feel of fingers running through his wavy hair. His number is 317-345-6864. Call or text him any time. ;)
-Charlie Dant
-Amanda Weber
-Sarah Huh
Sarah Huh? will be attending IU Bloomington in the fall. She is going to major in chemistry and go into pre-med to become a pediatrician. Currently Sarah is the 1st vice president of student council, in National Honor Society, Just Say No,and Fellowship of Christian Athletes. She was also the captain of her high school soccer team. Sarah plays travel soccer, sings for church, and paints soccer fields.
Project
Exposure to mercury is a widespread problem that affects many people all over the world. Most people ingest mercury through water sources. Mercury in water can arise from runoff from farms, chemical and industrial plants, household products in the trash, and sewage. Three types of mercury can adversely affect humans. Elemental, inorganic, and methyl mercury can all harm humans if ingested. Inorganic mercury is the most common form in drinking water and can cause kidney damage if enough is taken in. Methyl mercury is found in fish and humans can be exposed if they eat too much mercury-containing fish. Mercury ingestion can cause both acute and chronic symptoms.
We are using Saccharomyces cerevisiae yeast as a tool to detect mercury. In yeast, there are a number of transcription factors and genes that respond to oxidative stress and toxic metals. The yes associated protein (YAP) family is a family of transcription factors that is involved with oxidative stress regulation and redox homeostasis. They affect a number of genes, but we are focusing on GSH1 and GSH2. These genes are involved in the glutathione pathway. Glutathione is an antioxidant that protects the cell from oxidative stress.
In order to detect mercury, we are using several biological parts included in the BioBrick. The Kozak + mCherry translational unit is being used to give off a red fluorescent glow when the mercury is detected. In the plasmid, we will include the GSH2 promoter and the ADH1 terminator.
Notebook
1. Week 1 (02/16-02/20)
- Monday
Find background information on YAP1: YAP1 is a transcription activator involved in oxidative stress response and redox homeostasis made of 650 amino acids. YAP1 regulates the transcription of genes encoding antioxidant enzymes and components of the thiol-reducing pathways.
- Tuesday
Continue research on YAP1 and YAP family.
Read article on Gibson Assembly.
- Wednesday
Listen to lecture presentation on Gibson Assembly.
- Thursday
Continue research on YAP1 and YAP family.
- Friday
Wrap up research on YAP1 and write summary.
Begin to develop trial and error combinations for primer design
2. Week 2 (02/23-02/27)
- Monday
Begin primer design using the translational unit kozak+mcherry and the terminator ADH1.
- Tuesday
Use newly designed primer and attempt to construct it using Gibthon.
- Wednesday
Look into redox-center and interaction with disulfide bonds and cysteine residues.
Research on how mercury can affect individuals in the environment.
- Thursday
Research GSH1 and its enzyme gamma-glutamylcycteine synthetase: GSH1 (glutathione):GHS1 is an antioxidant that plays a role in the detoxification of oxidants and toxins from a cell.
Look into how YAP1 affects antioxidant enzymes such as gamma-glutamylcyctine synthetase.
- Friday
Continue Research on GSH1 and gamma-glutamylcyctein syntase.
3. Week 3 (02/30-03/03)
- Monday
Read article : The Role of Cysteine Residues as Redox-Sensitive Regulatory Switches by David Barford
- Tuesday
Continue reading article from Monday.
- Wednesday
Class discussion on IGEM project and progress in research.
- Thursday
Split into groups and begin working on the team wiki.
- Friday
Continue working on wiki notes and project overview.
4. Week 4 (03/05-03/09)
- Monday
Continue work on project overview.
- Tuesday
Presentation given by Dr. Goebal from IUPUI.
Take notes and ask questions
- Wednesday
Oxidation and ROS presentation by Forest.
- Thursday
Split into groups and continue writing for the wiki page.
Write about the YAP family, YAP1, GSH, and other components of the project.
- Friday
Type notes from notebooks into word document
Create Primer using Kozac+mCherry and ADH1; cut with ECOR1 and SPEC1.
5. Week 5 (03/12-03/16)
- Monday
Look over and present primer to group members.
- Tuesday
Discuss primer with class.
- Wednesday
Split into groups and create power points on specific topics i.e. YAP1
- Thursday
Continue working on power point.
- Friday
Practice power points and prepare for presentation to class.
6. Week 6 (03/19-03/23)
- Monday
Listen and take notes over YAP1 powerpoint.
- Tuesday
Listen and take notes over Glutathione presentation.
- Wednesday
Listen and take notes over Safety and Procedures presentation.
- Thursday
Add information from all presentations, including human impact, to team wiki page.
- Friday
Work on notebook.
7. Week 7 (03/26-03/30)
- Spring Break
8. Week 8 (04/02-04/06)
- Spring Break
9. Week 9 (04/09-04/13)
- Monday
Split into groups and research on the terminator and promoter while working on the Prezi and poster board.
- Tuesday
Continue working on Prezi and poster board.
- Wednesday
Welcome guest speaker from Community Hospital to discuss internships.
- Thursday
- Friday
10. Week 10
- Monday
- Tuesday
- Wednesday
- Thursday
- Friday
11. Week 11
- Monday
- Tuesday
- Wednesday
- Thursday
- Friday
Results/Conclusions
What did you achieve over the course of your semester?
Safety
Hazards:
POSSIBLE DANGERS TO THE ENVIRONMENT
Mercury is a metal that occurs naturally at low levels in rock, soil and water. Mercury is also released into the air, water and land .Most mercury pollution is released into the air and then falls directly into water bodies or onto land, where it can be washed into waterways. When mercury gets into water, bacteria can change it into a form called methylmercury, which is absorbed by tiny aquatic organisms. When fresh water and ocean fish eat those organisms, the mercury begins to build up in their bodies. When larger fish eat smaller fish, mercury can build up to high levels in the tissues of the big fish. Because it binds to the protein in fish muscles - the 'meat' of the fish - mercury cannot be removed by cooking or cleaning the fish.
PERSONAL INJURY DANGERS
Although our parts have a very little chance of harming our team members, there is still that very slight chance that a problem may arise. One of the parts that we work with is mercury, which, if come in contact with the human skin, can cause great harm and in the worst case scenario, death. Therefore, we will use the proper laboratory technique to avoid any and all consequences.
ASEPTIC TECHNIQUE:
Proper aseptic technique is essential to any lab experiment. It serves to create a barrier between microorganisms in the environment and the cell cultures that one is working with. Aseptic technique is comprised of four main components: good personal hygiene, a sterile work area, sterile reagents/media, and proper disposal/ clean up.
PERSONAL HYGIENE
• Be sure to wash hands your hands properly.
• Wear closed-toe shoes.
• Wear long pants that will cover your skin.
• If you have long hair, wear it up in a ponytail.
• Wear an apron/lab coat.
STERILE WORK AREA
• Choose an area to work in that is free from drafts and through traffic.
• Be sure the work area is uncluttered!
• Spread materials apart to keep from bumping things and to allow for proper air flow; keep things like pipets on the right/left hand side depending on which hand you use to hold it (If you’re right-handed, keep it on the right side).
• Clean lab table before the experiment!
• Use a 70% ethanol wash.
• CAUTION: Do not use ethanol in any area if a flame will be present, since it is highly flammable.
STERILE REAGENTS/MEDIA
• Wipe the outside of bottles, flasks, etc. with ethanol wash before placing them in the work environment.
• When opening a pipet, be sure not to let the tip touch anything.
• When opening bottles, don’t touch the inside of the cap; close back quickly!!
• Only use each pipet once!
PROPER DISPOSAL/CLEAN UP
• Wipe down all bottles/flasks again with ethanol once finished.
• Spray table with ethanol and wipe down once all materials are out of the work area.
• Contaminated waste goes in an autoclave bag! NOT THE TRASH CAN.
• If anything spills, mop it up immediately and then wipe with area with ethanol.
• Properly remove gloves.
Attributions
Who worked on what?
Human Practices
What impact does/will your project have on the public?
Fun!
What was your favorite team snack?? Have a picture of your team mascot?
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