DNA Club

DNA Club students have been working hard in the lab and have learned a lot about DNA related research and biotechnology. For many of them, this is their first taste of biology research and using biotechnology tools. 

Agar plate showing the bacterial host M. smegmatis with circular viral plaques (zones of death). Students successfully isolated a new bacteriophage that infects the host. These students will go on to purify the phage, name it and add to the phage library at the University of Pittsburgh.  Team includes Megan Roka, Nidhi Gadekar, Dhanvin Yajaman, Timi Oluwatimilehin, and Symrin Ranchod.

Jay Lothe and Lakshana Ramanan setting up PCR and counting first generation pillbugs for the Wolbachia pillbug feminization project.

DNA Club Project Updates 

Below are descriptions of the ongoing projects for 2021-2022. All projects started back in August and are in various stages of completion. These projects are all being done in house in our lab. All teams will produce a professional poster and short paper based on their results. These items will be on display in our annual symposium of research to be held in May.  

Posters and projects from the past 3 years can be found on the club website here: https://sites.google.com/view/dnaclub/home 

Phage Virus Team 

Our most ambitious and difficult project, the Phage Team is looking for bacteriophage viruses in soils and compost which might infect and kill a bacterium called Mycobacterium smegmatis (M. smeg). M. smeg is a close relative of tuberculosis and the goal is to identify and purify phages to be evaluated for use to combat tuberculosis and other bacterial diseases. Students have successfully isolated a phage from a compost soil sample and are in the process of purifying it. Once purified, they will extract DNA from the phage and prepare it for electron microscopy. These students will have the honor of naming this bacteriophage and contributing to the University of Pittsburgh phage library.  

US Ant Team 

This team’s goal is to identify the biodiversity of ants on IA campus and participate in the national US Ant biodiversity project. Students will identify ants through DNA analysis called “DNA Barcoding”. So far, the team has collected ants on campus from 6 zones and have extracted ant DNA, amplified a region of DNA used to identify species using PCR, and verified their extractions with a gel electrophoresis assay. They then sent their DNA for sequencing and their DNA sequences to establish ant taxonomy using bioinformatics and DNA barcoding. Track their progress HERE.  

Pillbug Team 

This team’s goal is to further an ongoing study detecting a bacterium (Wolbachia) that infects pillbugs and causes males to develop into females (feminization). Pregnant mothers were collected back in May and once they gave birth, they were preserved for analysis. Their offspring have been developing in culture. Students have begun extracting DNA from the mother pillbugs looking for the infectious Wolbachia bacteria. Students have also learned how to examine DNA sequence results to identify pillbug species and identify the strain of Wolbachia present. In addition, the offspring of each mother are being counted for ratios of male and female. Matching the mother’s Wolbachia status with the offspring ratios will allow us to evaluate the feminization effect. There are many strains of Wolbachia that infect arthropods. Our team is also running additional DNA tests to identify the specific strain infecting our pillbugs.  

Mosquito Team 

This team’s goal is to compare mosquito biodiversity between suburban yards with and without mosquito treatments and natural (non-suburban) surroundings. The Team collected mosquitos from August to mid-October from yards in Alpharetta and Dunwoody as well as Big Trees Forest Preserve and the Chattahoochee National Forest. They are now sorting mosquitos and DNA barcoding mosquitos to identify the species.  

B. apis Antifungal Team 

Bartonella apis (B. apis) is a bacterium that lives in the gut of honeybees and has been shown to kill fungi before they can infect a beehive. Our team is investigating how B. apis interacts with fungi in the hive environment including bee bodies, hive material, honey and nectar being brough into the hive by workers. The team has successfully worked at the hive, cultured fungus and is ready to test the fungus against bacteria. The team did a sequence of experiments, testing each fungal sample against the B. apis bacteria. In most trails, B. apis did protect against the cultured fungi. Students also extracted DNA from the fungi and DNA barcoded it to determine which type it is. We hope this may contribute to the science behind how B. apis bacteria protect hives from fungus.  

Honeybee Bacteria Team 

This team’s goal is to culture bacteria from various hive environments and products and investigate how they use energy. Students will do this by growing bacteria and then using a microplate treated with various metabolites to see which type of metabolism each bacteria type uses. On Friday, students will head to the hive to begin sampling and culturing their bacteria. Once cultured, it will be preserved until the metabolic experiments can be performed. Presently, the students are growing their control bacteria provided by the Newton Lab at Indiana University. They will conduct experiments to see which metabolites these bacteria prefer, whether they compete or share, and establish positive controls to evaluate their hive experiments in March.  

Pillbug Associated Fungi 

This team is looking at the specific fungi present in pillbug excrement. Pillbugs are decomposers and commonly live among fungus (also decomposers). They are curious to know which types of fungus are associated with pillbugs and how that relates to common fungi naturally present in the pillbug culture soil.  

Male Age Wasp Team  

The Wolbachia bacterial reproductive parasite affects arthropods differently. In the parasitoid wasp Nasonia vitripennis, Wolbachia will cause the death of offspring if an uninfected female mates with an infected male. Wolbachia is only inherited from mother or offspring, so in the wrong combination, Wolbachia prevents a dead end. This effect is called “cytoplasmic incompatibility” (CI). This team is testing to see if the age of the male at mating causes CI to breakdown and lose effect. They isolated male wasps and mated them with females in different Wolbachia combinations over a period of 6 days. They will count the offspring ratios looking for the strength of CI compared to controls.  

Temperature Wasp Team 

This is a Centennial High School team coming to IA to work. This team will also be looking at the effects of Wolbachia CI on Nasonia wasps. In this case, they will test the strength of CI offspring are developed at higher temperatures. In tropical areas of the world, Wolbachia induced CI is being used as a natural way to control disease spreading mosquitoes. Knowing how temperature affects CI gives insight to how well this control might work.