A Course-Based Undergraduate Research Experience (CURE) Using Transposon-directed insertion sequencing (TraDIS) to Identify Genes Required for Bacteriophage P1 Propagation in Escherichia coli

Here, we describe the design and implementation of a multi-week course-based undergraduate research experience (CURE) embedded within a third-year Microbial Genetics course. In this CURE, students construct a high-density Tn5 transposon insertion library in Escherichia coli, challenge the library with bacteriophage P1, and use Transposon-Directed Insertion-site Sequencing (TraDIS) coupled with statistical analysis (edgeR) to identify genes required for phage propagation.

Students perform transposome electroporation, phage infection assays, genomic DNA extraction, high-throughput sequencing data processing in Galaxy, differential insertion analysis, visualization in Artemis, and Gene Ontology enrichment analysis. The project integrates wet-lab microbiology, molecular genetics, and bioinformatics while modeling an authentic forward genetic screen. This CURE emphasizes experimental design, genome-wide functional analysis, statistical reasoning, and interpretation of large-scale sequencing data.