MICROBIO 565 Laboratory in Molecular Genetics

The demand for authentic research experience at UMass greatly outstrips supply. One way to bridge this gap is to offer course-based undergraduate research experiences, or CUREs. Microbio 565 is a CURE that focuses on the cell envelope of mycobacteria. These organisms are technically Gram-positive but have an unusual, complex envelope that in some ways is more reminiscent of Gram-negatives. They grow weirdly, expanding their envelope preferentially from one end of the cell (instead of evenly along its entire length á la Escherichia coli and other well-studied species). In Fall 2018 we will build on work done by Ph.D. student Emily Melzer with the help of then-undergraduate Caralyn Sein. They have shown that the protein DivIVA initiates and stabilizes polar growth in Mycobacterium smegmatis by organizing cell wall peptidoglycan synthesis. We will repeat some of their experiments, then extend this work by asking 1) whether DivIVA organizes peptidoglycan assembly by localizing MurG, a cytoplasmic enzyme required for precursor synthesis and 2) what part(s) of the DivIVA protein contribute to its function. Guided by the literature, in the last, post-Thanksgiving third of the class, we will perform experiments of the students’ choosing. In Microbio 565 we will use genetic depletion and site-directed mutagenesis along with methods from cell and chemical biology to address a real research question. Neither the professor nor the TA knows the answer! This means that some things may not work as hypothesized or may not work at all. The class will puzzle together to figure out whether the hypothesis needs adjusting or whether something went wrong with the execution of the experiment. If the latter, we map out a troubleshooting plan. Besides gaining hands-on experience with modern genetic techniques, students learn how to learn from failure, an essential skill in bench science. 
Semester(s) offered: 
MICROBIO 312 with a C or higher