Course code:
457B2
Course name:
Protein Engineering

Academic year:

2022/2023.

Attendance requirements:

There are no requirements.

ECTS:

4

Study level:

graduate academic studies

Study program:

Biochemistry: 1. year, winter semester, elective (E52B2) course

Teacher:

Radivoje M. Prodanovi, Ph.D.
full professor, Faculty of Chemistry, Studentski trg 12-16, Beograd

Assistant:

Karla . Ili uri, Ph.D.
assistant with PhD, Faculty of Chemistry, Studentski trg 12-16, Beograd

Hours of instruction:

Weekly: two hours of lectures + two hours of labwork (2+0+2)

Goals:

Students should get knowledge about state of the art of protein engineering and design.

Outcome:

Know how of theoretical and experimental methods for protein engineering and design.

Teaching methods:

Lectures, labwork, semmester papers.

Extracurricular activities:

Coursebooks:

  1. . : , - , 2019.
  2. Sheldon J. Park, Jennifer R. Cochran: Protein Engineering and Design, CRC Press 2010.
  3. S. Brakmann, K. Johnsson: Directed Molecular Evolution of Proteins, WILEY-VCH 2002.
  4. Selected review papers

Additional material:

  Course activities and grading method

Lectures:

10 points (2 hours a week)

Syllabus:

  1. Theories of protein evolution and in silico protein design.
  2. In vitro protein evolution - principles and troubleshooting.
  3. Surface display of proteins in Protein engineering.
  4. In vitro translation of proteins in Protein engineering.
  5. Gene library preparation - gene duplication, circular permutations, gene fusion, exon shuffling, mutations, consensus mutagenesis.
  6. High throughput screening methods in Protein engineering - principles and troubleshooting.
  7. Bioinformatics in Protein engineering.

Labwork:

20 points (2 hours a week)

Syllabus:

  1. Preparation of gene libraries by saturation mutagenesis, recombination and error prone PCR.
  2. Transformation of yeast S. cerevisie and expression of gene library on the surface of yeast cells.
  3. High throughput screening of gene libraries in microtiter plates.
  4. High throughput screening of gene libraries using in vitro compartmentalization and flow cytometry.

Semester papers:

20 points

Oral exam:

50 points