Faculty of Chemistry and Chemical Technology, Akereva 5, Ljubljana

- 1-

30-1016-en
1 (2)

1. SUBJECT: RECOMBINANT DNA TECHNOLOGY

Code: 30-1016
Credits: 5.0
Form of teaching: 60; Lectures 60 hours
Programme: University Undergraduate Programme Biochemistry

2. OBJECTIVES

Applicative aspects of recombinant DNA technology are emphasised. Techniques required for
generation of recombinant DNA molecules and preparation of recombinant proteins are
presented, as well as several applications in molecular biology, medicine, agriculture and
forensics.

3. SYLLABUS

A: History. Legislation in Slovenia and EU. Laboratory organisms. Safety aspects.
B: DNA replication and transcription; protein biosynthesis (resume). Electrophoretic
techniques. Enzymes in recombinant DNA technology.
C: Vectors. Reporter genes. Cell transformation. cDNA synthesis and preparation of libraries.
D: DNA synthesis in vitro. DNA sequencing. Polynucleotide chain reaction (PCR).
Hybridisation. Labelling of specific probes. DNA-microarrays.
E: Expression systems general. Expression in prokaryotes: vectors, fusions, optimisation of
production; targeted localisation, solubility, stability, refolding.
F: Expression in yeasts and filamentous fungi. Expression in cultured animal and human cells.
G: Approaches to mutagenesis. Protein engineering. Surface display. Yeast two-hybrid
systems.
H: Forensic genetics. DNA-diagnostics. DNA technology in systematics and ecology.
Production of recombinant reagents.
I: Bioremediation and biomass utilisation. Recombinant bacteria in agriculture. Recombinant
DNA in plant systems.
J : Genetic engineering of plants: methodology and applications. Genetically modified
organisms. Transgenic organisms; knockout technology. Gene silencing by RNAi.
K: Stem cells. Cloning of mammals. Human genome project and determination of other
genomes.
L: Genomics - proteomics. Pharmacogenomics. Recombinant DNA technology in medicine.
Gene therapy.
M: Patenting in recombinant DNA technology. Ethics and public acceptance.
N: New approaches in recombinant DNA technology.

4. PREREQUISITES

Biochemistry, Molecular genetics, Microbiology.

5. RECOMMENDED TEXTBOOKS

MAIN TEXTBOOK:
B.R. Glick, J .J . Pasternak: Molecular Biotechnology, Principles and applications of
recombinant DNA, 3
rd
edition, ASM Press, Washington, 2003.

ADDITIONAL LITERATURE:
S. Primrose, R. Twyman, B. Old: Principles of Gene Manipulation , 6
th
edition, Blackwell
Science, Oxford, 2001.
http://bio.ijs.si/katedra/TrDNA1.html (in Slovenian).
http://bio.ijs.si/katedra/rDNA-Technology.html (English page with basic data only).

6. FORM OF ASSESSMENT

Written and oral examination.


PREPARED BY: Marko Dolinar

DATE: 20.10.2003

30-1016-en
2 (2)