RECOMBINANT DNA TECHNOLOGIES
Academic year and teacher
If you can't find the course description that you're looking for in the above list,
please see the following instructions >>
- Versione italiana
- Academic year
- 2017/2018
- Teacher
- DARIO BALESTRA
- Credits
- 6
- Didactic period
- Secondo Semestre
- SSD
- BIO/11
Training objectives
- The Recombinant Technologies represent a discipline in which all of those methods and experimental approaches that are used inside of a Molecular Biology laboratory in order to engineer and manipulate DNA and proteins , in order to study their role and / or function and transfer them into the genome of other cells by controlling the incorporation and expression are addressed.
The main objective of the course is to provide students with the molecular basis of the methods and applications most used within a laboratory of Molecular Biology and provide the basis to apply the gained knowledge.
Knowledge and understanding
Students will:
- know the proper terminology
- know the molecular basis of the main laboratory techniques
Applyed knowledge and understanding
Students will:
- know how to properly use the scientific terminology
- know how the methods of a Molecular Biology Laboratory work
- know how to design a Molecular Biology experiment
- know how to analyze and choice the proper plasmid vector Prerequisites
- No formal propedeuticity is required. However, being a multi-disciplinary course, knowledge of physic and chemistry is necessary. Moreover, a strong background in Biochemistry and Molecular Biology is mandatory.
Course programme
- The course will be split into 3 main parts:
in the first part ( 20 hours ) , after a brief introduction to recombinant technologies and their evolution over time, arguments concerning the cloning of a DNA fragment will be developed. In particular will be addressed:
- PCR and its variants
- cut and paste of DNA molecules, restriction enzymes, ligase, TOPO-cloning
- DNA sequencing, pyrosequencing
- DNA fragments analysis, electrophoresis and DNA quantification by qPCR
- plasmid vectors, phages, cosmids, BAC, PAC, YAC
In the second part (12 hours), approaches involved in the creation and analysis of genomic libraries and expression libraries will be discussed. In particular:
- design and construction cDNA and genomic libraries
- selection and screening strategies
- analysis of cloned DNA
- transformation and transfection of genes in prokaryotic and eukaryotic cells, including plants
- Reporter genes
- yeast to study eukaryotic genes
In the last section (16 hours), methods, approaches and strategies used to engineer proteins, gene expression and creation of transgenic animals will be discussed. In particular:
- fusion proteins
- vectors for gene therapy
- RNAinterference
- molecular evolution in vitro, DNA shuffling
- Cre-Lox approach
- inducible promoter Didactic methods
- The course is structured in frontal lectures for a total of 48 hours. Lessons are provided on a weekly basis by taking advantage of power-point slides and of the classical backboard. The last 4 hours of the course are devoted to summarize the main contents of the program.
Learning assessment procedures
- The aim of the exam is to test the level of knowledge and deepening of the topics of the course program and the reasoning skills developed by the student on issues related to biochemistry. The assessment is expressed in thirtieths (minimum grade 18).
With the exception of students manifesting problems, the exam is written, and consists of 6 open questions on the main topics of the course. Each response value can score up to 5. In any case, the exam is not considered passed whether the student scores =2 in two or more questions. Reference texts
- Being a multi-disciplinary course, there are not texts containing all topics
Specific topics can be further developed in the following texts.
- Teacher’s handouts
- "INGEGNERIA GENETICA" Sandy Primrose, Richard Twyman, Bob Old, Zanichelli.
- "DNA RICOMBINANTE"J.D. Watson et al., Zanichelli"BIOTECNOLOGIA MOLECOLARE"B.R. Glick, J.J. Pasternak, Zanichelli
- "BIOTECNOLOGIA MOLECOLARE"B.R. Glick, J.J. Pasternak, Zanichelli
