Purpose: The purpose of this lab was to determine how many of the collected Bay Area insects have the Wolbachia bacteria in their DNA
What is Wolbachia?
Wolbachia is a bacteria infecting arthropods and nematode worms that can change the sex of the host as well as kill their offspring
Why is Wolbachia being studied? What did you discover in your research?
Studies have shown that Wolbachia can be used to stop the mosquitos that spread diseases like zika and dengue and also finding ways to end this virus can be useful in curing diseases like Lymphatic filariasis (elephantiasis) and Onchocerciasis (river blindness.)
What kinds of relationships does Wolbachia have with other organisms?
Wolbachia is parasitic to other organisms as this bacteria benefits at the expense of the other organism Specifically, Wolbachia is an endosymbiotic parasite which means that it lives inside the host cell.
I was going to list out all the steps, but that would be a very long list! So, instead, I will summarize. First, we all brought in bugs from our houses or our surrounding areas and froze them in alcohol. Next, we mashed up the abdomen of the bug, and if your bug was a certain size, you had to cut it to make sure you just get the reproductive parts of the insect! I needed to cut mine because I caught (my dad caught) a moth and this bug was pretty big! After the abdomen was all smushed in our microfuge tubes, we had to separate the DNA from the pellet (the stuff we do not need). We did this by centrifuging many times and adding different liquids like NaCl and Lysis Buffer which bind together to give us a clear DNA sample. After going through multiple steps, it was time for the Polymerase Chain Reaction (PCR) machine which replicates the DNA thus allowing the electrophoresis gels to be clear. Also on the gel electrophoresis, we included a positive control and a ladder both of which help us determine what is DNA and what is Wolbachia so thus we know whether or not the insect had been infected with Wolbachia bacteria.
Personally, my moth was not infected by Wolbachia and had a visible DNA on the gel! Out of the 11 girls who performed this experiment, there were only 4 insects infected with Wolbachia and all of them came from the Redwood city/ San Carlos area. A coincidence? I think not.
The experience of this lab was much easier and more fun than the other labs because I was more prepared being that I understand the topic and work more. Maybe it was the fact that this was my last lab and I had all the knowledge of my other labs in the back of my mind, or maybe I was just less stressed. Regardless, this was definitely my favorite lab being that I 100% understand what was going on. I understood all the tools we were using like the centrifuge and electrophoreses, even though occasionally I had micropipette troubles.
My partner Gina and I created a presentation of DNA technology and pharmaceutical products! We chose this topic because both of our parents have experience at Genentech, a company making great advancements in personalized medicine in oncology! We discussed how insulin is made, how personalized medicine works, and an ethical question regarding personalized medicine! Check out our powerpoint and the script google doc to follow along and leave your thoughts below!
Medical Applications: Pharmaceutical Products SCRIPT!!!
The pharmaceutical industry is significantly benefitted by advances in DNA technology and genetic research. Advancements can be applied to the development of useful drugs.
Defined as the use of living things or parts by means of recombinant DNA technology in order to create or modify drugs
Products are created using organic chemistry or biotechnology
Biopharmaceutical = a biological macromolecule or cellular component used as a pharmaceutical.
Pharmaceutical companies are producing molecules made by recombinant DNA to treat various diseases
According to the National Center for Biotechnology Information (NCBI), “a majority of therapeutic drugs in the current market are bioformulations, such as antibodies, nucleic acid products and vaccines”
insulin for diabetes
interferon for viral infections
Interferon = a group of signaling proteins made and released by host cells in response to the presence of several pathogens, such as viruses, bacteria, parasites, and also tumor cells
Production of human hormones and proteins with therapeutic uses
Enhanced weight loss
Producing safer vaccines
Now being produced commercially from cloned genes
Example & DNA Technology
Recombinant DNA is a technology that makes it possible to insert a human gene into the genetic material of a common bacterium, thus the “recombinant” (re-combined) microorganism can now produce the protein coded by the human gene. Like with insulin!
People with diabetes have a problem with how their body produces or responds to insulin, therefore scientists use recombinant dna to produce insulin medicine for diabetics to take.
First the scientists build the human insulin gene in the lab and then they remove a loop of bacterial dna which is known as plasmid
Next they insert the human insulin gene into the plasmid and then researchers return the plasmid to the bacteria and put the recombinant bacteria into a large fermentation tank
Then the recombinant bacteria use the gene to produce human insulin which scientists then harvest from the bacteria and purify to make medicine for people
Oncology, which is the study and treatment of tumors and cancers, has become increasingly personalized at Genentech because medicine that works for one person, may not work for another!
Researchers at Genentech stated that the realization that human diseases and many cancers are heterogeneous, which means they result from different molecular drivers that fuel the disease, was the key driver
Two women with breast cancer whose disease looks similar under the microscope can have VERY different diseases at the molecular level and thus need different treatments
The more information they have, the better they can understand the role of different mutations in each disease, develop biomarkers and identify the right medicine for EACH patient
Many Biomarkers + multiple targeted medicines = endless combinations designed to treat many different types of cancer
The major concerns with pharmaceutical product biotechnology:
the current “EpiPen Scandal”
EpiPen is priced in the $600s but CVS will be stocking a generic Adrenaclick which come in a two-pack for $109.99
Very expensive, created with biologic material as opposed to synthetic material thus making the medicine more expensive
Only certain populations can afford this medicine as insurance companies who pay for the treatments will make the patient pay increasing amounts
Thus the question is created, should personalized medicine only be for those who can afford the treatment, or should the care be readily available to everyone in need. If so, at what cost?
Our last project before our semester final was to pick a disease or disorder that interests you, and find out ALL you can about it with the caveat that you must be able to discern where the exact gene that causes the mutation is located. My partner Michelle and I decided to choose Down Syndrome because this syndrome is huge, affecting many people in America. However, Down Syndrome is not a single gene…Down Syndrome is the presence of a whole extra chromosome! To find out more, check out our voice thread and feel free to leave feedback!
Purpose: To observe and determine the phases of the cell life cycle by identifying stages of mitosis in a root tip.
Introduction: Cells at the tip of a growing plant are constantly dividing, allowing the root to grow. In Mitosis, a cell doubles it’s chromosomes and then divides into two identical copies of the original cell. These cells divide independently, thus the cells in the root tip are at different stages of cell division. These stages include interphase, the pre-phase where a cell is just a dark mass as the cell doubles it’s unorganized chromosomes and prepares for division. Prophase, where the chromosomes are visible in a microscope as they condense. Centrioles move to opposite sides of the cell. In Metaphase, the spindle fibers move the chromosomes to the equator of the cell and centrioles are at opposite ends. Next comes Anaphase where the sister chromatids are pulled apart to opposite poles. Finally in Telophase, two daughter cells form and each have a full set of chromosomes as well as a nucleus.
Onion root-tip slides
Set onion root-tip slide on the set-up microscope stage
Focus the microscope to clearly view the onion root under the 40x lens, once viewed, change the lens to 400x and focus finely until the cells are clear
Count vertically the longest line of cells, then count horizontally the longest row of cells. Multiply these two numbers, the result is the total number of cells in the stage.
Identify the cells in the phases of Telophase, Anaphase, Metaphase and Prophase, in this order. Record the number of each phase in a table
Calculate the total number of cells in each phase of the previous step, then subtract that number from the total number of cells in the stage. The result is the number of cells in Interphase, record in the table.
Looking at two more onion root-tips, repeat steps 2-5 and record.
Data Table (Personal)
Data Table (Class Averages)
The majority of the cells were in Interphase
Percentages in each stage
The evidence that shows that mitosis is a continuous process is that each phase is hard to differentiate which showcases how the process is a flow and not definitive.
In each cell observed, there are 4X chromosomes being that Interphase doubles the original 2X chromosomes.
After Meiosis, each sex cell would have 1X chromosome being that each daughter cell (2X) divides in half.
In the zygotes produced, there would be 2X chromosomes.
Based on the data recorded from the class, the cells are in Interphase 81% of the time which is completely reasonable being that the cells need to function as opposed to just reproducing.
Meiosis goes a little something like this. The process starts with Interphase, which happens before Meiosis even begins. In interphase, the cell grows, copies all of its chromosomes, and then prepares for division. Following this, Prophase 1 begins and the chromosomes condense as well as the nuclear envelope breaks down. Also in this phase, crossing over occurs! After condensing, the chromosomes pair up and each chromosome aligns with its homologue partner so that the two match up at corresponding positions along their full length so they can share part of their DNA. Next comes Metaphase 1 where the pairs of homologous chromosomes move to the equator of the cell. Then during Anaphase 1, the homologous chromosomes move to opposite poles of the same cell. In the end of Meiosis 1, the chromosomes gather at the poles of the cells and the cytoplasm divides. That phase is known as Telophase and cytokinesis. Next comes Prophase 2, where a new spindle forms around the chromosomes. Following the same pattern comes Metaphase 2, the chromosomes line up at the equator. In Anaphase 2, the centromeres divide and the chromatids move to the opposite poles of the cells. Lastly, in Telophase 2 and Cytokinesis, a nuclear envelope forms around each set of chromosomes and the cytoplasm divides leaving 4 daughter cells.
I worked with Veronica and our process went pretty smoothly. Our school work time did not go as perfectly as we had hoped, being that we had to rerecord a lot of our data which set us back time wise. Luckily we were able to finish 90% of our recording at school and then we met at Veronica’s house on Sunday to finish up and put everything together. I think making a video really helped us to fully understand Meiosis because comparing our knowledge before and after the video, we have grown immensely.
Merzlyak1*, Mark N., Olga B. Chivkunova1, and Alexei E. Solovchenko1 And. “Mark N. Merzlyak.” Light Absorption by Anthocyanins in Juvenile, Stressed, and Senescing Leaves. N.p., 12 Aug. 2008. Web. 03 Nov. 2016.
This season, the 2016 AP Bio class created Breakthrough Junior Challenge videos (https://breakthroughjuniorchallenge.org) in an attempt to not only become experts on exciting topics, but to compete for a chance to be recognized for our work! My video is on adrenaline, because I notice it’s effects so much in my day to day life. Sometimes, the effects are small. Like when I have a performance or competition for cheer and I notice my hands are shaking and my heart is beating out of my chest. Or sometimes the effects are bigger like when a highway police officer is hidden on the road as I am racing to get home and for a second I think I am about to be dragged away to prison for life!!! It happens to all of us. We shake, our hearts race, our breath escapes us, once I even got an immediate headache! But why does this happen? How does our body do this? What are our cells doing? Well watch my video and you will find out.