2014/2015

2016/2017

DM270

BIO/13 (APPLIED BIOLOGY)

DEPARTMENT OF BIOLOGY AND BIOTECHNOLOGY "LAZZARO SPALLANZANI"

BIOLOGICAL SCIENCES

PERCORSO COMUNE

3°

1st semester (01/10/2016 - 14/01/2017)

6

72 lesson hours

italian

ORAL TEST

BINDA CLAUDIA (titolare) - 2 ECTS
CANOBBIO ILARIA - 3 ECTS
FORNERIS FEDERICO - 1 ECTS

basic knowledge of molecular biology and biochemistry

The aim of the course is to provide the students with the basic theoretical and practical tools required to work in a laboratory and, in particular, to learn the methods to carry out experiments of DNA manipulation and of purification, biochemical characterization and crystallization of proteins.

First part: bacterial genomic DNA extraction; restriction digestion of genomic and plasmid DNA; DNA gel electrophoresis; generation of restriction map; cloning of pyruvate kinase cDNA in an expression vector.
Second part: preparation of buffer solution and pH measurement; usage of chromatographic techniques to purify pyruvate kinase; protein gel electrophoresis; enzymatic assays.
Third part: crystallization experiments of pyruvate kinase and of lysozime by different techniques; analysis of the results and phase diagram determination; computational structural biology: computer practicals using softwares for determination and analysis of three-dimensional structures of pyruvate kinase and other biological macromolecules.

Short lectures to introduce the topics, followed by practicals.

Books used for the courses of Biochemistry and Molecular Biology.

Report on experiments carried out during the practicals and test to evaluate the learning of the basic concepts.

Students are required to bring a lab coat to wear during the experiments.