Student Research Assistant
Job Description
A science major exposed to hands-on experience beyond the classroom will increase her analytical skills and prepare her for a graduate track or a research assistant position.
The student will gain confidence and independence by generating a research project and carrying it to completion. Even if the career path is different than an academic track, critical thinking will help her in a health-related profession.
Year 1 ~ Responsibility/Function
The student will become familiar with the laboratory environment, safety regulations and ethical behavior. She will get trained on maintaining the cultures with Entamoeba spp cells, prepare culture media, sterilize old cultures/medial, order reagents and supplies. She will be trained in molecular biology techniques including DNA isolation, cloning, gene expression, PCR, cell growth, protein purification and inhibition. The student will be exposed and participate in weekly laboratory discussions on scientific literature related to the research and will present a discussion at the end of the Spring or Summer 2021.
She will start as soon as approved (Spring 2021) and work during Fall 2021. At the end of the Spring 2022 the student will present a poster with her proposal for a research project to work during summer and Fall-Spring of year 2.
My research combines structural/biochemical analyses of anaerobic enzymes, ecological adaptations of these enzymes and cell behavioral strategies to develop improved strategies to manage amebiasis. Entamoeba histolytica, the etiological agent of amebiasis, causes infections of 50 million people every year and 100 thousand fatalities worldwide. Despite the epidemiological importance of E. histolytica, there are currently no ideal methods to prevent infection in areas without appropriate sanitation. We are interested in furthering our knowledge to develop improved/non-toxic strategies to manage amebiasis in humans.
The student will work in one of two projects:
1. Essential enzymes for the survival of Entamoeba and inhibiting strategies to manage the parasistes. Anaerobic metabolism / Enzyme Kinetics (molecular microbiology, genetics, biochemistry).
2. Entamoeba Clone-recognition Experiments: Evolution, Ecology and Implications to Health. Clusters of Entamoeba trophozoites from different phylogenetic lineages show distinctive rate of aggregation, density of cells per cluster, and distance between clusters. This project aims at the identification/ characterization of the discrimination/recognition genes in Entamoeba. By understanding the biological/evolutionary basis of kin-discrimination/recognition we could modify our approaches in the management of disease.
Year 2 ~ Responsibility/Function
The student will start her own project.
If she picks project 1, she will extract DNA from Entamoeba spp. cells, design primers for the ADHE gene isolation, isolate the gene by PCR, clone it in a plasmid, transform bacteria and express the protein. Because it is a time-consuming project, this will take about a year. If time remains the student will start the purification and biochemical analysis (see year 3). The student will present posters in campus (Academic Showcase Spring 2022) and departmental and college events.
If she picks project 2, she will explore the mechanisms of discrimination that Entamoeba spp varieties use to aggregate with members of their own group and distance from members of a different group. We have proposed that the identification of cell-signals secreted by the amebas that are responsible for discrimination should be the next steps. This would be the first multi-clone characterization of Entamoeba spp. aggregative behavior, and cell- signaling secretion in the context of clone-recognition. Protists allow us to study cell–cell recognition from ecological and evolutionary perspectives. Modern protistan lineages can be central to studies about the origins and evolution of multicellularity. She will use fluorescent dyes to mark different populations and design physical spaces to study motility and aggregative behavior of amebae populations.
Year 3
Responsibility/Function
If the student picks project 1, the students will purify the protein (ADHE), analyze the function biochemically by determining substrate and cofactor binding properties (kinetics). The student will verify expression and purification by gel analysis and antibody specificity binding. This process will take a year. During this time the student will present posters in campus (Academic Showcase SP) and in departmental, college events. The student will also have to write a final paper with the results and conclusions. If everything goes well, the student will be an author in the submitted scientific manuscript.
If the student picks project 2, the student will work on detecting and confirming the chemical molecules involved in the signaling between individual and clustered cells. We have shown that Entamoeba discriminate between one another depending on the relatedness of the taxa. Previous experiments have provided strong evidence to support the presence of kin recognition and cell signaling. Signaling and aggregative behavior could impact pathogenicity and virulence. Based on preliminary data, we have proposed that analogous signaling pathways to single cell eukaryotes may exist in Entamoeba. Methods were developed to help determine the structure and composition of the chemical signaling molecule, including the use of conditioned and unconditioned media to cause aggregation. The student will examine the resulting chemical signaling molecule(s) using proteomic analysis and antisense molecular mechanisms to block the expressions of the molecules. If molecules are identified and blocked, the results will help develop better strategies for the identification and phylogenetic classification of different varieties of Entamoeba. These strategies can also be used in designing better approaches to manage amebiasis.
*Students are prohibited from driving during work-study employment with the limited exception of authorized work-study driver positions.
** All employees, including student employees, are required to participate in a short online training that will prepare you with the new protocols for working on campus. Please click on the following link to complete your training:http://rwu.co1.qualtrics.com/jfe/form/SV_a4aBCUkcc7Vl7QF