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551-1263-00L 3 Credits
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Gene and Protein Technology

Lecturers & Examiners: Dr. Ned Mantei, Prof. em. Dr. Hans-Martin Fischer, Prof. Dr. Dario Neri
VVZ CR n/a

Last Updated: 2026-02-05 14:54:57

Objective

Part 1: (H.M. Fischer) In the first part basic principles and methods that are widely used in molecular genetics are discussed. These include: Isolation, analyis and cloning of nucleic acids. Cloning in bacteria and yeast. Construction of libraries. Polymerase chain reaction and selected applications. Directed mutagenesis. Analysis of gene structure and expression. Part 2: (N. Mantei) Gene cloning, whole genomes, transgenic organisms, recombinant protein production, recent developments. Part 3: (D. Neri) Antibody Engineering, Chemical Modification of Proteins, Measurement of affinity constants, Proteomics

Content

Part 1: In the first part basic principles and methods that are widely used in molecular genetics are discussed. These include: Isolation, analyis and cloning of nucleic acids. Cloning in bacteria and yeast. Construction of libraries. Polymerase chain reaction and selected applications. Directed mutagenesis. Analysis of gene structure and expression. Part 2: Isolation of cDNA clones - how to find the right one? Differential expression analysis - microarrays, differential display, etc. Dealing with whole genomes - microsatellites, modern genetic maps, positional cloning, ESTs, FISH. Transgenic and "knockout" organisms. Production of "recombinant" proteins. Recent developments with promise for the future. Part 3: I) Antibody phage technology: the antibody molecule V genes, CDRs, basics of antibody engineering principles of phage display phagemid and phage vectors antibody libraries phage display selection methodologies other phage libraries (peptides, globular proteins, enzymes) alternative screening/selection methodologies intrabodies, plantibodies robotization by-passing antibody technology II) Proteins: chemical modification and biomolecular interactions chemical modifications of proteins labeling proteins with photoactive molecules homo- and hetero-dimerisation of proteins radioactive labeling of proteins (alpha, beta and gamma emitters) kinetic association and dissociation constants affinity constant: definition and its experimental measurement III) Proteomics and related technologies Immobilised pH gradients 2D-PAGE Prefractionation/Prefiltration (plasma) 2D-DIGE Mass-spectrometry and protein identification (tryptic digests, MALDI-TOF, LC-MS-MS etc.) Isotope-coded affinity tags (ICAT) Functional Proteomics (probing serine hydrolases, identifying protein-protein interactions) In vivo Proteomics Protein arrays, antibody arrays Biosensor technologies (BIAcore)

Resources

Lecture Notes

Part 1: PDF files will be available on a weekly basis athttp://www.micro.biol.ethz.ch/under "Lecture Scripts" (username and password will be giben)Part 2: PDF files will be available on a weekly basis athttp://www.cell.biol.ethz.ch/teaching/Gentechnology(username and password will be given at one of the early lectures)Part 3:1) Skript [http://www.pharma.ethz.ch/files/course_materials]2) Trail, P.A., Willner, D., Lasch, S.J., Henderson, A.J., Hofstead, S., Casazza, A.M.,Firestone, R.A., Hellström, I. and Hellström, K.E. (1993). Cure of xenografted humancarcinomas by BR96-doxorubicin immunoconjugates. Science, 261, 212-215.3) Mann, M., Hendrickson, R.C., Pandey, A. (2001) Analysis of proteins and proteomesby mass spectrometry. Annu. Rev. Biochem., 70, 437-4734) Adam GC, Sorensen EJ, Cravatt BF. (2002) Chemical strategies for functional proteomics.Mol. Cell. Proteomics. 1, 781-790.Recommended further reading1) Winter, G., Griffiths, A.D., Hawkins, R.E. and Hoogenboom, H.R. (1994) Making antibodiesby phage display technology. Annu. Rev. Immunol. 12, 433-455.2) Gygi. S.P., Rist, B., Gerber, S.A., Turecek, F., Gelb, M.H., Aebersold, R. (1999).Quantitative analysis of complex protein mixtures using isotope-coded affinity tags.Nature Biotechnology, 17, 994-999.3) Huber, L. (2003) Is proteomics heading in the wrong direction? Nature Reviews MolecularCell Biology, 4, 74-80.4) De Wildt, R.M.T., Mundy, C. R., Gorick, B.D., Tomlinson, I.M. (2000) Antibodyarrays for high-throughput screening of antibody-antigen interactions. NatureBiotechnology, 18, 989-994.5) Carter, P. (2001) Improving the efficacy of antibody-based cancer therapies. NatureReviews Cancer, 1, 118-29.6) Abbott, A. (2002) Betting on tomorrow's chips. Nature, 415, 112-114.

Literature

1. Dale, J.W. and von Schantz, M. 20002. From genes to genomes. Concepts and applications of DNA technology. John Wiley & sons Ltd. Chichester, UK 2. Reece, R.J. 2004. Analysis of genes and genomes. John Wiley & sons Ltd. Chichester, UK 3. Gibson, G. and Muse, S.V. 2002. A primer of genome science. Sinauer Associates, Inc. Publishers, Sunderland Mass., USA

General Information

Language
English
Frequency
Yearly recurring

Examination

Type
session examination
Mode
oral 30 minutes

Course Components

Type Title Time & Place Hours
lecture with exercise Gene and Protein Technology
  • Mon 15:15-17:00 (HG E 7)
  • Wed 13:15-14:00 (HG D 7.1)
3 h weekly

Offered In