Our Progress So Far Group 2: Hannah Pastore, Grace Ruh, Czavier Tan, - - PowerPoint PPT Presentation

Presentation #2

Our Progress So Far

Group 2: Hannah Pastore, Grace Ruh, Czavier Tan, Alyssa Levy, Lunide Sylne and Javier Nodarse

Agenda

1) TEP1 2) What do we want to do? 3) Our hypothesis

TEP 1

• Human tumor suppressor gene

○ Protein facilitates the repair of shortened telomeres ○ Not the only gene involved

• A short telomere = bad

○ Aging ○ Cancer ○ Death

What do we want to do?

• Take out TEP1
• Shortened telomeres can lead to many different results

○ Our purpose is to find out the specific role TEP1 has on telomere length maintenance

• So does TEP1 help prevent:

○ Aging? ○ Cancer? ○ Cell death?

Our Hypothesis

• Modified yeast cells will exhibit some symptoms

synonymous with shortened telomeres

• Based on observations, we should get a better

understanding of what TEP1 does for a telomere. For example:

○ Rapid growth = cancer ○ Slowed reproduction = aging ○ Increased death = cell death

Our Experiment

• 1. Making our PCR solution

a. Finding primers b. Combining all of our solutions

• 2. Checking the results of PCR

a. Gel electrophoresis

• 3. Yeast: from test tube to agar plate

a. Preventing contamination

• 4. Centrifuging & Incubating

Creating Our PCR Solution

• What is PCR?

○

The process of heating and cooling a sample of DNA in such a way that it is denatured, annealed, and extended to amplify certain sequences of genetic code

• Primers:

○ IDT website - find a sequence of 20-30 nucleotide bases at the beginning and end of the gene sequence that would react around 62°C, the temperature at which PCR would function

• Antibiotics

○ “Genetic markers” ○ Those with the TEP1 gene would die while those without the gene would survive - way to effectively observe what occurs in absence of the gene

• Process

○ Added 8 components ○ Worked with the other lab groups to create a solution we could all use prior to including our gene specific components

Did Our PCR Work?

• Results of gel electrophoresis showed that our PCR was not

effective

• Why?

○ Human error (i.e. spilling, not measuring the correct amounts) NO :(

Time to Grow Our Yeast

Control Group (Day 1)

Suspended yeast cells were combined with growth solutıon in four separate tubes and plated on an agar plate with specific arrangements to ensure identification of contamination (hopefully none) later

Experimental Group (Day 2)

Use of centrıfuge and ıncubatıon periods to separate yeast components, unravel DNA, and ultimately transform them to remove TEP1 gene; plated on agar plates and distributed with glass beads

Has Our Hypothesis Changed?

• Previously, we thought TEP1 and telomeres were only associated with cancer

○ Now, we know that telomeres control a variety of factors ○

Ex) TEP1 in Malaria

■ Limits parasite development ■ Essential for parasite killing

○ Ex) TEP1 and Fertility

■ Damaged telomeres cannot support a zygote

• Our first hypothesis was that our cells would have a higher growth rate and

experience shorter lifespan if they didn’t have the TEP1 gene. However, due to readings and observations, we think that the cells will continue to grow normally and not experience an early death.