Course Description
Topics in this course include plasmid construction, CRISPR-Cas9 mediated mutagenesis, plant transformations, PCR, recombinant protein expression and purification, protein engineering, protein structure prediction with AlphaFold, protein-ligand docking and other bioinformatics analyses. This course introduces student to current recombinant, molecular and biochemical techniques and covers some of the basic ethical issues in recombinant DNA Technology. The adopted techniques are important for us to study the biological functions of proteins from various sequenced genomes. These techniques will include, analysis of CRISPR-Cas9 system for plants, Agrobacterium-mediated transformation of Arabidopsis, genomic DNA extraction from Arabidopsis, phenotypic analysis of Arabidopsis gene deletion lines, construction of bacteria protein expression vectors, design of functional mutants, analysis of protein expression and purification from E. coli and protein crystallization. Students will be introduced to some common bioinformatics tools, including AlphaFold and Pymol for protein structure prediction and analysis, Blast and ClustalW for sequence analysis. Enzymes of the shikimate pathway will be the subjects for our studies. The shikimate pathway plays an important role in the biosynthesis of secondary metabolites and plant hormones. In Arabidopsis, perturbations of the pathway result in distinct and visible phenotypes, some of which will be the focus of this course. The laboratories are complemented with lectures to discuss theoretical approaches and ethical issues pertinent to the laboratory materials.
Prerequisite
BIO230H1 / BIO255H1
Recommended Preparation
BIO251H1 or higher level plant biology course,
BCH311H1 / CSB349H1 / MGY311Y1 taken concurrently
Ancillary Fees
This course has a lab fee of $50.
Lab coat and safety glasses are required for the in-person lab component and the approximate cost is $25. Students are responsible for purchasing these items.
Lecturer(s)
Prof. D. Christendat
dinesh.christendat@utoronto.ca
Prof. E. Nambara
eiji.nambara@utoronto.ca
Contact Hours
24L, 36P
Required Text(s)/Readings
No required textbook.
Students will be provided with primary literature as required readings.
Evaluation (Subject to change)
This is a typical mechanism for evaluation from the previous years of offering. Please note that the official course evaluation (marking scheme) will be provided at the beginning of the semester
Lab Report 1 (Module 1)
Progress report
Lab Report 2
Quizzes
Lab Participation
Final Exam
Last updated on June 21st, 2024