Bioremediation

Environmental Microbiology and Biotechnological Remediation:
Environmental biotechnology utilizes microorganisms to improve the environmental quality. These improvements include the treatment of contaminated waters and wastewaters, clean up of industrial waste streams, and remediation of soils contaminated with hazardous and toxic chemicals.
This course will introduce the students to the scientific principles of microbiological treatment technologies to clean up contaminated environments and to the basic DNA engineering techniques to manipulate microorganisms for these purposes. Moreover, the students will acquire experience on molecular methods /techniques used to monitor the presence of contaminants by the construction of a microbial biosensor. The course will consist of lecture-based teaching to provide background information on key concepts on bioremediation and in a laboratory-based section where the students will construct and use a microbial biosensor.
The lecture-based section will include the following topics: (1) fundamentals of environmental microbiology, (2) principles of bioremediation of organic contaminants and toxic molecules, (3) engineering strategies for bioremediation and biosensors construction.

Laboratory section

Day 1: Extraction of genomic DNA from suitable bacterial strain and quantification.
Day 2: Amplification of regulative gene and promoter by PCR, check on agarose gel and excision from agarose gel. Enzymatic digestion of fragment DNA.
Day 3: Plasmid DNA extraction (Midiprep) and purification. Enzymatic digestion.
Day 4: DNA fragment/plasmid excision from agarose gel, set up of the ligation mixture and overnight incubation.
Day 5: Competent Escherichia coli cells preparation and transformation. Overnight incubation.
Day 6 (second week): Plasmid preparation (Miniprep) from transformants and screening by restriction analysis.
Day 7: Growth of the reporter E. coli and control strains in the presence of different xenobiotic compounds and samples collection.
Day 8: β-galactosidase assay. Conclusions.

Laboratory of Cell Biology Cell culture techniques are commonly used in cell biology, pharmacology and toxicology. Starting from basic approaches and considerations, this course aims to provide the theoretical and practical skills needed for designing cell culture-based experiments. It will also provide an overview of the technologies available for manipulating the gene expression of cultured cells and integrating this approach into a research project. The lecture-based section will include the following topics: (1) fundamentals of cell culture; (2) novel cell culture technologies; (3) fluorescence-based techniques; (4) methods to assess cell proliferation, viability and survival; (5) methods to assess cell adhesion and migration; (6) transient and stable gene overexpression; (7) other technologies of genetic manipulation. The practical section will include demonstrations on cell culture and manipulation.