Protein engineering and Synthetic Biology - MB151P131

• Title: Protein engineering and Synthetic Biology
• Czech title: Proteinove inzenyrstvi a synteticka biologie
• Guaranteed by: Department of Cell Biology (31-151)
• Faculty: Faculty of Science
• Actual: from 2023 to 2023
• Semester: winter
• E-Credits: 3
• Examination process: winter s.:combined
• Hours per week, examination: winter s.:2/0, Ex [HT]
• Capacity: unlimited
• Min. number of students: unlimited
• 4EU+: no
• Virtual mobility / capacity: no
• State of the course: taught
• Language: English
• Note: enabled for web enrollment; the course is taught as cyclical
• Guarantor: Mgr. Klára Hlouchová, Ph.D.; RNDr. Cyril Bařinka, Ph.D.
• Teacher(s): RNDr. Cyril Bařinka, Ph.D.; Mgr. Klára Hlouchová, Ph.D.

Requirements to the exam

Last update: Mgr. Klára Hlouchová, Ph.D. (17.02.2021)

2 assignments / homeworks

final grade: 25+25% for assignments, 50% final exam

 

Syllabus

Last update: Mgr. Klára Hlouchová, Ph.D. (07.10.2020)

1. Introduction (why protein engineering?, antibodies as an example of engineered proteins), DNA sources and manipulation

2. Expression plasmids, cloning/mutagenesis techniques (restriction sites, Gateway, Gibson assembly, In fusion)

3. Heterologous expression systems (prokaryotic, yeast, insect, mammalian) - transfection, cultivation, disruption

4. Basics of amino acid/protein structure, protein purification techniques I (affinity tags, ion exchange, hydrophobic, size-exclusion)

5. Protein purification techniques II, protein analysis (purity, concentration, dispersity, function, structure)

6. Repetition/back-up

7. Cell-free synthesis

8. Creating Diversity / Libraries

epPCR, degenerate oligo design, gene shuffling, library preparation

9. Expanded / Alternative Genetic Code

codon reassignment

10. Screen vs Selection towards new function

display techniques (Phage, ribosomal, mRNA/cDNA), compartments

11. Minimal cells

genome reduction

12. Synth Bio

metabolic engineering; gene circuits