Molecular Parasitology
Swati Agrawal, University of Mary Washington
In Spring 2021, we piloted a mini-CURE where student groups from University of Mary Washington and Georgia State University collaboratively completed research projects as part of a research-intensive course on Molecular Parasitology. The benefits of this approach were immediately obvious as students interacted across institutions, learned from each other's disciplinary expertise while informing their own research with data collected by their collaborators.

The HICA project
Kathleen Cornely, Providence College
In this CURE, inspired by the work of Hoffmann, et al., students prepare mutant Haemophilus influenzae carbonic anhydrase (HICA) proteins. Using PyMOL to visualize the three-dimensional structure of the HICA protein, students choose one or more surface amino acid residues to mutate to histidine residues in order to create a surface histidine cluster that will allow the mutant protein to bind to a nickel affinity column. Using site-directed mutagenesis, recombinant plasmids are constructed and are then used to transform an E. coli expression vector. The mutant HICA protein is overexpressed, cells are lysed, and students load the cell lysate onto Ni-NTA columns and determine the imidazole concentration required to elute the mutant protein. The construction of a library of mutant proteins will allow the development of a general method in which specific surface histidine residues of any protein can be mutated in order to facilitate affinity purification. The Haemophilus influenzae bacterium described herein is a respiratory pathogen that causes meningitis (in its encapsulated form) and mucosal infections such as otitis media, sinusitis and conjunctivitis (in its unencapsulated form). A recent study showed that the carbonic anhydrase enzyme is absolutely required for pathogenesis. Furthermore, expression of the HICA enzyme allows the pathogen to survive in host immune cells (Langereis, et al.). These observations make the study of HICA itself particularly attractive, in addition to the overall goal of contributing to a body of work that will allow the minimal histidine character required for nickel affinity to be ascertained.

Exploring eukaryotic protein structure and post-translational modifications.
Erica Jacobs, St. John's University-New York
This CURE will provide opportunity for students to think and act as researchers by using computational, biochemical, and bioanalytical techniques to examine tick antigen proteins. The CURE is designed as a lab for upper-level students who are taking or have taken a one-semester introductory biochemistry course, but two semesters would be even better. It could also be adapted for cell/molecular biology or (bio) analytical chemistry instrumentational analysis labs. It has been taught for classes ranging from 12-24 students. Ticks are notorious vectors of viral, protozoan, and bacterial diseases, including Lyme disease. While an anti-vector vaccine capable of protecting people from diseases transmitted by a particular tick species is an alluring goal, only one such anti-tick vaccine is currently available. This vaccine targets Bm86, a protein from the midgut of Rhipicephalus microplus, a cattle tick. Not only does the vaccine limit parasitism of the cattle by ticks, data suggests that it can also prevent transmission of tick-borne diseases including bovine anaplasmosis and babesiosis. However, similar vaccination approaches have not succeeded thus far against ticks that transmit diseases to humans, and little is known about the antibody response to the antigen, or about the protein itself. Since the protein's structure and function are unknown, the research goal of this CURE is to purify Bm86 using an insect cell/baculovirus expression system and characterize it, including domain structure and post-translational modifications (glycosylation sites). There are homologs to Bm86 in every sequenced tick species examined, and future CUREs will characterize some of the homologs including those in Ixodes scapularis, the tick that is mainly responsible for transmitting Lyme in the eastern US, and Haemaphysalis longicornis, the Asian longhorned tick, a newly-discovered invasive species in the area that also has significant disease-transmitting potential. By understanding the structure and post-translational modifications of this protein, we hope to gain a better understanding of how to make effective anti-tick vaccines, including those for humans, that may prevent transmission of Lyme disease. Importantly, the basic parameters of this CURE can be used to examine other proteins besides tick antigens. For example, during the pandemic, the CURE pivoted from the tick antigen to the SARS-CoV-2 nucleocapsid protein, which was also expressed in an insect cell system. Instead of characterizing glycosylation sites, we characterized phosphorylation sites. It's therefore possible to use this same framework for many different eukaryotic proteins that may be of research interest.

Biomass conversion into highly useful chemicals
SAPNA JAIN, Alabama State University
This is CURE based course that aims at bridging the gap between theoretical knowledge in chemistry and its practical applications at solving real-world problems. It gives students an opportunity to construct and synthesize their knowledge and skills by learning to apply theoretical knowledge to practice by the laboratory research. The purpose of this course is to acquaint students with the fundamental concepts of chemistry, synthetic methods and techniques. The emphasis will be on novel catalysts synthesis and evaluating their activity towards biomass conversion to liquid fuel and useful chemicals. Students will design synthesize, deduce identities of the biomass conversion products from chemical and spectral clues, and predict reaction products.

Synthesis of the Intermediate of a Catalytic Reaction: An NHC-Stabilized, First-Row Transition Metal Complex
Meng Zhou, Lawrence Technological University
The advanced synthesis laboratory course object allows students to study the synthesis, purification, and characterizations of a new diamagnetic organometallic complex of a first-row transition metal. The air-stable complex is stabilized by an N-heterocyclic carbene spectator ligand. It also bears an actor ligand and therefore, is potentially a reactive intermediate of a catalytic reaction. The synthesis of a reactive intermediate is the key to elucidate the mechanism of catalysis. The instructor chooses the first-row transition metal and the actor ligand based on his or her interests. The CURE starts from an NHC-ligated complex that does not bear this actor ligand but is otherwise similar. In our CURE, an anion ligand-replacement reaction was used to install the actor ligand, but an instructor may choose other approaches. The students will evaluate their results by standard spectroscopic analyses using UV-vis, FT-IR, and proton NMR (60 MHz or above) analysis.

Characterizing the Aging Process Using Caenorhabditis elegans and Reverse Genetics
Joslyn Mills, Bridgewater State University
Using gene silencing (RNAi) in the nemotode C. elegans, students will identify genetic modifiers of proteins with roles in aging by reverse genetics. Specifically, students will analyze the effect of knocking down genes on the level of aging-related proteins tagged with fluorophores (GFP, RFP, etc.). Each group of students will use function-specific RNAi libraries (transcription factors, kinases, etc) already established in our lab. Furthermore, students will evaluate the effect of genetic modifiers on proteostasis and lifespan. In addition to becoming familiar with C. elegans work and appreciating the use of model organisms, the students will master microscopy, genetic crosses, gene silencing, and molecular and biochemical readout assays such as qPCR and immunoblotting.

An Arabidopsis Mutant Screen CURE for a Cell and Molecular Biology Laboratory Course
Jinjie Liu, Michigan State University
This CURE is designed from a crucial component of a chloroplast lipid signaling research project and has been implemented for a cell and molecular biology laboratory course at Michigan State University. The research laboratory generated an engineered plant line producing a lipid-derived plant hormone and mutagenized this line. The research question is "what transporters or receptors are involved in the hormone signaling transduction or perception processes?". Students form research hypotheses based on the research model, design experiments, perform experiments, collect and analyze data, make scientific arguments, and share their findings with the learning community. Specifically, the students culture the mutagenized plant population and select the desired mutant phenotypes, followed by genotyping the mutants and characterizing the mutants by basic biochemical approaches. Mathematics is also integrated into the course design. As the students studied the relevant genetic, molecular and biochemical concepts during this CURE, they use the core idea of information flow and data they generate in the lab to make claims about their mutant plants and support these claims with evidence and reasoning.

Population & Community Ecology
Cascade Sorte, University of California-Irvine
Students in a Population and Community Ecology class participate in coastal marine research focused on understanding factors determining population sizes and community interactions, particularly in the context of species that appear to be shifting their ranges with climate change. Students participate in all aspects of the research from making observations and collecting data in the field to defining questions, stating hypothesis, designing and completing statistical analysis, and interpreting and presenting results. The outcomes are a research proposal, research paper, and poster presentation. All are intended to be at a level appropriate for use as a writing sample or presentation at undergraduate conferences. Results are incorporated into the ongoing research project led by the course instructor and graduate student teaching assistant.

Organismal Form and Function Lab
Christopher Oufiero, Towson University
Invertebrates use movement of their bodies and structures in diverse ways to interact with their environment. This includes general locomotion (e.g., walking, jumping, flying) to specific forms of locomotion (e.g., gliding on water), using limbs to acquire food (e.g., raptorial forelegs in the praying mantis) and using structures to communicate (e.g., cricket calls). These movements have been the focus of bioinspiration studies to understand how these small organisms, with compact nervous systems, are able to achieve their movements. Given the diversity of invertebrates and the lack of information on the variation in their movements, the goals of this course are to understand the variation in invertebrate movement and explore the factors that may affect that variation. In this course, students have the opportunity to develop and test their own research hypotheses associated with variation in the movement of invertebrates. Using high-speed cameras, students are instructed on filming techniques to quantify animal movement, the use of the R programming language to obtain basic kinematics of movement and analyze their data, and the process of science from hypothesis formation to presentation of results. Research questions change each iteration based upon the hypotheses students develop, but the same instructional material and skillsets (e.g., quantifying animal movement) are consistently used. Results from each student group are presented during a departmental wide poster symposium and can be written up for publication, where applicable.

Design2Data
Ashley Vater, University of California-Davis
The D2D program is centered around an undergraduate-friendly protocol workflow that follows the design-build-test-learn engineering framework. This protocol has served as the scaffold for a successful undergraduate training program and has been further developed into courses that range from a 10-week freshman seminar to a year-long, upper-division molecular biology course. The overarching research goal of this CURE probes the current predictive limitations of protein-modeling software by functionally characterizing single amino acid mutants in a robust model system. The most interesting outcomes of this project are dependent on large datasets, and, as such, the project is optimal for multi-institutional collaborations.