title: Investigation of the human disease osteogenesis imperfecta: a research-based introduction to concepts and skills in biomolecular analysis.
abstract: A blended approach encompassing problem-based learning (PBL) and structured inquiry was used in this laboratory exercise based on the congenital disease Osteogenesis imperfecta (OI), to introduce commonly used techniques in biomolecular analysis within a clinical context. During a series of PBL sessions students were presented with several scenarios involving a 2 year old child, who had experienced numerous fractures. Key learning goals related to both the theory and practical aspects of the course, covering biomolecular analysis and functional genomics, were identified in successive PBL sessions. The laboratory exercises were conducted in 3 hour blocks over six weeks, focused firstly on protein analysis, followed by nucleic acids. Students isolated collagen from normal and OI affected fibroblast cultures. Analysis by SDS-PAGE demonstrated α1 and α2 of collagen Type I chains at approximately 95 kDa and 92 kDa, respectively. Subtle differences in protein mobility between the control and OI samples were observed by some students, but most considered it inconclusive as a diagnostic tool. The nucleic acid module involved isolation of RNA from OI affected fibroblasts. The RNA was reverse transcribed and used as template to amplify a 354 bp COL1A1 fragment. Students were provided with the sequence of the OI affected COL1A1 PCR product aligned with the normal COL1A1 sequence, allowing identification of the mutation, as the substitution of Arg for Gly(976) of the triple helical region. Our experience with student cohorts over several years is that presentation of this laboratory exercise within a relevant clinical context, and the opportunity for active engagement with the experimental procedures via PBL sessions, supported the learning of basic theory and practical techniques of biomolecular analysis.
authors: School of Biomedical Sciences and Pharmacy, University of Newcastle NSW Australia. karen.mate@newcastle.edu.au
authors: Karen
authors: K
authors: Mate
authors: Alistair
authors: A
authors: Sim
authors: Judith
authors: J
authors: Weidenhofer
authors: Liz
authors: L
authors: Milward
authors: Judith
authors: J
authors: Scott
mesh_terms: D001483:Base Sequence; D035843:Biomedical Research / Q000193:education*; D002478:Cells, Cultured; D002675:Child, Preschool; D024042:Collagen Type I / Q000737:chemistry* / Q000235:genetics*; D000090602:Collagen Type I, alpha 1 Chain; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010013:Osteogenesis Imperfecta / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018794:Problem-Based Learning; D012313:RNA / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D018076:DNA, Complementary; D012313:RNA