Team:WarrenCentral WCC IN

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	Team WarrenCentral_WCC_IN

Official Team Profile

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 and plays tennis for recreation.

-Cheyenne Harvey

-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 Funny 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

My name is Sarah Huh? And I will attending IU Bloomington in the fall. I am going to major in chemistry and go into pre-med to become a pediatrician. I am currently the 1st vice president of my student council, in National Honor Society, Just Say No, Fellowship of Christian Athletes. I was also the captain of my high school soccer team. I also play travel soccer, sing for my church, and paint 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  Begin power point on YAP1 - Thursday  Continue working on power point. - Friday  Practice power point about YAP1 with Myka, who made a power point o the YAP family.

6. Week 6 (03/19-03/23) - Monday  Present YAP1 power point - 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.

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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