CH-319 / 4 credits

Teacher: Hovius Rudolf

Language: English

Withdrawal: It is not allowed to withdraw from this subject after the registration deadline.


Summary

A 7-week long (4+8 h) experiment where you plan and construct a fluorescent sensor protein starting from DNA bricks. The protein will be expressed in and purified from E.coli, characterized by biochemical and spectroscopic methods. Your report will be in the style of a scientific paper or poster

Content

i) Molecular biology: DNA fragment isolation by PCR, DNA restriction and ligation, plasmid purification and sequence analysis, agarose gel electrophoresis.

 

ii) Biochemistry: Bacterial protein expression, protein purification by affinity chromatography, analysis by SDS-PAGE and spectroscopy, fluorescent labelling

 

iii) Biophysics: Fluorescence spectroscopy, enzyme kinetics or molecular interactions.

Keywords

Molecular biology - cloning

 

Protein expression & purification

 

Bio-chemical and bio-physical chracterisation

 

Sensor proteins

 

Reporting

Learning Prerequisites

Required courses

Admission to the TP is conditional on the successful completion of 2 of the 3 courses: CH-210 Biochimie, CH-313 Chemical biology, CH-311 Macromolecular structure and interactions

 

Recommended courses

Molecular and cellular biophysics I (CH-311)

Chemical Biology (CH-313)

Important concepts to start the course

genetic engineering & DNA manipulation; protein synthesis; DNA & protein analysis; absorbance and fluorescence spectrometry; enzymology, molecular interactions

Learning Outcomes

  • Design cloning strategy
  • Produce a scientific report and high-quality lab journal
  • Integrate Good laboratory behavior and wet lab practice
  • Assess / Evaluate your data critically
  • Produce a purified expressed protein
  • Analyze proteins and DNA
  • Characterize sensor function
  • Use common sense and logical deduction
  • Interpret protein structure data
  • Design cloning strategy
  • Produce a scientific report and high-quality lab journal
  • Integrate Good laboratory behavior and wet lab practice
  • Assess / Evaluate Evaluate your data critically
  • Produce a purified expressed protein
  • Analyze proteins and DNA
  • Characterize sensor function
  • Interpret protein structure data

Transversal skills

  • Collect data.
  • Continue to work through difficulties or initial failure to find optimal solutions.
  • Write a scientific or technical report.
  • Plan and carry out activities in a way which makes optimal use of available time and other resources.

Teaching methods

Students prepare and discuss experimental approach

 

Laboratory experimentation

 

Discussion of experimental progress and results

 

Reporting on scientific level

Topiocal presentations

Expected student activities

Good theoretical preparation & planning of lab work before doing experiments; skillful execution of experiments, being organized & keeping a lab book; thorough analysis of results, writting a scientific-grade report; respecting security rules and fellow students.

Assessment methods

valuation of preparation and planning

 

Evaluation of experimental execution, including good laboratory behaviour

 

Evaluation of comprehension through discussion & written questions

 

Evaluation of report: including structure, data treatment and presentation, critical attitude , comparison to scientific literature

 


Supervision

Office hours Yes
Assistants Yes
Others

Resources

Bibliography

Bibliography

 

TP manual

 

Biochemistry & Biophysics text books

 

ApE; a plasmid editor free software

 

Methods, Structures, and other useful info via Moodle

Notes/Handbook

Manual

Websites

Moodle Link

Prerequisite for

big plus for Msc courses and for semester or diploma projects in chemical biology or biophysics

 


In the programs

  • Semester: Spring
  • Exam form: During the semester (summer session)
  • Subject examined: Experimental biochemistry and biophysics
  • Practical work: 6 Hour(s) per week x 14 weeks
  • Type: mandatory
  • Semester: Spring
  • Exam form: During the semester (summer session)
  • Subject examined: Experimental biochemistry and biophysics
  • Practical work: 6 Hour(s) per week x 14 weeks
  • Type: optional

Reference week

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