n.BA.BT.PrMB1BC2.22HS (Molecular Biology 1 and Biochemistry 2 Practical) 
Module: Molecular Biology 1 and Biochemistry 2 Practical
This information was generated on: 19 April 2024
No.
n.BA.BT.PrMB1BC2.22HS
Title
Molecular Biology 1 and Biochemistry 2 Practical
Credits
3

Description

Version: 2.0 start 01 August 2023

 

Study Programme Biotechnology
Regulations Applicable RPO, 29 January 2008, School of Life Sciences and Facility Management Academic Regulations, 15 Dec. 2009, Annex for the Bachelor of Biotechnology degree programme
Module Type  
X Compulsory Module    Elective Module    Optional Module
Planned Semester 3rd Semester
Module Coordinator Martin Sievers
Telephone / E-Mail +41 (0)58 934 57 16 / martin.sievers@zhaw.ch
Lecturer(s),
Speaker(s),
Associate(s)		 Andrea Baier, Tobias Wermelinger, David Frasson
Entrance Requirements Molecular Biology 1, Organic Chemistry and Biochemistry 1
Learning Outcomes and Competencies As part of the practicals, an enzyme is produced recombinantly from a bacterium in Escherichia coli BL21(DE3). The recombinant protein is purified, visualised in SDS PAGE and the specific activity of the recombinant protein determined. Molecular biological techniques such as PCR, cloning with the aid of restriction enzymes, production of competent E. coli cells, DNA ligation, transformation, plasmid isolation, DNA sequencing, protein purification and enzyme activity determination is learned and specifically applied in the course.
Students will then be able to independently design and implement cloning strategies for the recombinant production of proteins in Escherichia coli.
Module Content The module focuses on the following practical steps, leading from gene to product:
 
  • Isolation of DNA from a bacterium
  • PCR: Amplification of the CoDing Sequence (CDS) for the enzyme to be cloned, starting from the DNA isolated. 
  • Restriction enzyme digestion of the DNA vector and PCR product
  • Ligation of the PCR product into a DNA expression vector 
  • Transformation into E. coli NEB 5-alpha
  • Plasmid isolation, DNA sequencing of the insert, sequence evaluation
  • Transformation into the expression strain E. coli BL21(DE3)
  • Induction, lysis, protein purification, dialysis
  • SDS-PAGE, Western blot for process control
  • Determination of the specific activity of the recombinant enzyme
  • Enzyme kinetics
Follow-up Modules Molecular Biology 2 and Immunology 1
Methods of Instruction  Practicals
Digital Resources Guide to Practicals
Lesson Structure / Workload  
 Contact Hours 42
 Guided Self-Study -
 Independent Self-Study 48
 Total Workload 90
Classroom Attendance -
Assessment
Written exam at the end of the semester 100%

If there is a low number of participants, the lecturer may change the form of a repeat examination after consultation with the head of the study programme: e.g. an oral examination can be used to replace a written one. Please report any changes to the form of examinations by e-mail to pruefungsadmin.lsfm@zhaw.ch and Cc. Head of study programme.
Language of Instruction  German
Comments

Nicola A. Burgess-Brown, Heterologous Gene Expression in E.coli: Methods and Protocols, Humana Press, 2018

Jens Kurreck, Joachim W. Engels, Friedrich Lottspeich, Bioanalytik, Springer Spektrum, 2022

 

Note

Course: Molecular Biology 1 and Biochemistry 2 Practical
No.
n.BA.BT.PrMB1BC2.22HS.P
Title
Molecular Biology 1 and Biochemistry 2 Practical

Note

  • No module description is available in the system for the cut-off date of 19 April 2024.