Collagen IV Assembly: Production of a Recombinant Construct for Mechanistic Studies

Basement membranes, specialized extracellular matrix, provide mechanical support for epithelial cells and shape cell behavior by interacting with cell receptors. Collagen IV, the predominant component of basement membrane, comprises six genetically distinct alpha chains (α1 to α6) that assemble to form protomers, which associate to form hexamers, in the scaffold. Non-assembly due to mutations within alpha chains give rise to kidney diseases. Yet, the mechanism of assembly is poorly understood. Preliminary data in our lab indicated that amino acid 78 of collagen IV α2 NC1 (non-collagenous) domains may play a role in a chloride-dependent switch that supports hexamer formation. We hypothesized that this particular amino acid is critical for hexamer assembly. To this end, we sought to generate a mutated construct of collagen IV α2 truncated protomer, TP(D78A). We anticipate this construct will be a critical tool in understanding the mechanism of collagen IV α2 domain assembly. Using molecular biology techniques, we introduced a point mutation in TP(D78A) and identified optimal conditions for protein expression. We generated PCR products of expected molecular weight and sent them for sequence verification. We then investigated cell lines for optimum protein expression to later look for hexamer formation. We found that COS-7 cells as compared to HEK-293 cells were better for expression of our wild-type construct WT(TPD78). In conclusion, we produced candidate TP(D78A) constructs for sequencing and determined COS-7 cells were better for expression of WT(TP(D78).These results are significant in producing a tool to investigate the mechanism of collagen IV α2 NC1 domain assembly

Undergraduate Biology Education Research Gordon Research Conference: A Meeting Report

The 2019 Undergraduate Biology Education Research Gordon Research Conference (UBER GRC), titled “Achieving Widespread Improvement in Undergraduate Education,” brought together a diverse group of researchers and practitioners working to identify, promote, and understand widespread adoption of evidence-based teaching, learning, and success strategies in undergraduate biology. Graduate students and postdocs had the additional opportunity to present and discuss research during a Gordon Research Seminar (GRS) that preceded the GRC. This report provides a broad overview of the UBER GRC and GRS and highlights major themes that cut across invited talks, poster presentations, and informal discussions. Such themes include the importance of working in teams at multiple levels to achieve instructional improvement, the potential to use big data and analytics to inform instructional change, the need to customize change initiatives, and the importance of psychosocial supports in improving undergraduate student well-being and academic success. The report also discusses the future of the UBER GRC as an established meeting and describes aspects of the conference that make it unique, both in terms of facilitating dissemination of research and providing a welcoming environment for conferees