Molecular Cell Biology and Biotechnological Applications (2020/2021)



Course code
4S008240
Credits
6
Coordinator
Linda Avesani
Academic sector
AGR/07 - AGRICULTURAL GENETICS
Language of instruction
Italian
Teaching is organised as follows:
Activity Credits Period Academic staff Timetable
Biologia cellulare molecolare teoria 2 II semestre Flavia Guzzo

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Biologia cellulare molecolare laboratorio 1 II semestre Mauro Commisso

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Bioreattori vegetali teoria 2 II semestre Linda Avesani

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Bioreattori vegetali laboratorio 1 II semestre Linda Avesani

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Learning outcomes

Learning objectives Frontal Lessons -recognize the complexity of the eukaryotic cell and list the sites of production and destination of the proteins with particular emphasis on the plant cell -define the “targeting” and “sorting” concepts; - describe the processes of vesicles formation and fusion and the mechanism of cargo selection; - identify the different paths of the proteins to their final destination; - distinguish the different information necessary for targeting a protein along the default pathway and along the other paths; -follow the post-transductional modifications of proteins along the secretion pathway focusing on quality control processes; - acquire an appropriate scientific language; - apply the acquired knowledge to eukaryotic cell engineering. Laboratory - prepare fresh plant samples for observation with standard and confocal microscopes; -use various types of fluorescence microscopes: conventional, inverted, stereomicroscope, confocal; - choose reporter proteins and fluorescent probes to mark various sub-cellular compartments based on the complementarity of emission spectra and available facilities (types of microscopes and filters); -observe and interpret the intercellular localization of fluorescent reporter proteins in various compartments through fluorescence and confocal microscopy, using specific probes to identify the compartments; - Acquire images under fluorescence and confocal microscopes and analyze them. Bioreactors Module Learning objectives Frontal Lessons -to define the concept of molecular farming; -to distinguish between stable and transient expression of recombinant proteins in plant systems; -to describe the major applications of molecular farming, including the production of nanomaterials, antibodies and vaccines; -to discuss the various strategies to optimise the production of recombinant proteins in plant systems, including the one based on human glyco-engineering, and their fields of application; -to evaluate the impact of different downstream processing strategies in the production system; -to plan production systems for molecules with medical and industrial applications using plants. Laboratory -to design genetic constructs with different fluorescent reporter proteins that accumulate in different sub-cellular compartments; -to prepare constructs with different fluorescent reporter proteins that accumulate in different sub-cellular compartments; -to express transiently those reporter proteins in the leaves of the model host plant species Nicotiana benthamiana; -to measure the yields of the expressed proteins using semi-quantitative western blotting and to compare the efficacy of each single approach.