ADVANCED MOLECULAR BIOLOGY
Learning outcomes of the course unit
The objective of the course is to bring the student into a deeper knowledge of:
- buffer systems with a special care for the ones in the blood;
- relevance and applications of the osmotic pressure;- use of the modern molecular biology techniques, emphasizing heterologous expression and protein engineering.
Course contents summary
Relevance of pH and osmotic pressure in the homeostasis process.
Ionic equilibria of poliyprotic acids and their salts in aqueous solution. Acid-base behaviour of amino acids in aqueous solution. Titration of amino acids. pH-buffer systems in the blood. Preparation of buffer solutions.
Origin and significance of the osmotic pressure. The van’t Hoff coefficient. Calculating osmolarity. Preparation of isotonic solutions.
Reactions and characterization of carbohydrates.
DNA manipulation techniques
In vitro mutagenesis: random and site-specific mutagenesis, and PCR-mediated mutagenesis reaction.
Production of recombinant proteins
Heterologous expressions in yeast cells. Fusion proteins; the pMAL system in Escherichia coli.
Applications of genetic engineering techniques
Analysis of DNA polymorphisms. Diagnosis of genetic diseases.
Searching the nucleotide database, query by DNA sequence.
All the study materials and supporting web sites are provided by the teachers.
Frontal lectures (aided by presentations and animations files) and practicals.