Teaching code

4S008240

Credits

6

Coordinator

Linda Avesani

Language

Italian

Scientific Disciplinary Sector (SSD)

AGR/07 - AGRICULTURAL GENETICS

Moodle Seminars 0

The teaching is organized as follows:

Learning objectives

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.

Prerequisites and basic notions

Basic knowledge of molecular biology

Program

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MM: Biologia cellulare molecolare teoria
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-Few synthesis sites, many final destinations: protein trafficking in the eukaryotic cell, with particular focus to the compartments and organelles of the plant cell. - Protein trafficking along the secretion pathway (RE, GA, TGN, PM, V); targeting and sorting. - The mechanisms of vesicular traffic: formation and fusion of vesicles, their control, cargo selection. - Protein modifications along the secretion pathway and quality control.
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MM: Biologia cellulare molecolare laboratorio
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- Kohler illumination. - Fluorescence microscopy. - Staining of plant tissues with fluorescent dyes to highlight cellular compartments. - Staining of in vitro plant cell cultures with fluorescent dyes to highlight cellular compartments. - Observation and identification of cellular compartment highlighted with protein fluorescent probes.
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MM: Bioreattori vegetali teoria
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Molecular cell Biology: -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.
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MM: Bioreattori vegetali laboratorio
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Didactic methods

Frontal lessons

Learning assessment procedures

Oral exam for each module

Evaluation criteria

Knowledge of course topics

Criteria for the composition of the final grade

Oral exam grade

Exam language

italiano