Course Identification

From DNA cloning to protein expression, purification and structural analysis
20203462

Lecturers and Teaching Assistants

Dr. Shira Albeck, Dr. Yoav Peleg, Dr. Orly Dym, Dr. Tamar Unger, Dr. Yael Fridmann-Sirkis
N/A

Course Schedule and Location

2020
Second Semester
Dates: 10th of May till 27th of May (12th of May vacation) Hours: 1.5 h daily around 14:00,
10/05/2020
27/05/2020
30

Field of Study, Course Type and Credit Points

Life Sciences: Laboratory; Elective; Regular; 1.00 points

Comments

Dates: 10th of May till 27th of May (12th of May vacation)
Hours: 1.5 h daily around 14:00

Prerequisites

No

Restrictions

50

Language of Instruction

English

Attendance and participation

Obligatory

Grade Type

Pass / Fail

Grade Breakdown (in %)

10%
90%
participation obligatory

Evaluation Type

Final assignment

Scheduled date 1

N/A
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-
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Estimated Weekly Independent Workload (in hours)

N/A

Syllabus

This theoretical course is intended to provide an overview on various aspects of protein engineering including molecular DNA manipulations, recombinant protein expression in various hosts, protein purification & characterisation and structural analysis. 

 

The following lectures will be given:

  • Introduction to DNA manipulations & recombinant proteins
  • Strategies in DNA cloning  
  • DNA mutagenesis  
  • Protein expression in bacterial cells 
  • Yeast expression system 
  • Protein expression in Baculovirus-infected insect cells and mammalian cells 
  • Introduction to Proteins & Purification 
  • Purification of proteins by FPLC  
  • Handling purified proteins 
  • Refolding Inclusion Bodies (IB) 
  • Correlation between Sequence, Function & Structure 
  • Protein Crystallization & Crystallography 
  • Introduction to PyMoll
  • Analysis of protein-protein interaction using MST and Biacore

Learning Outcomes

Upon successful completion of this course students should be able to:

Gain basic knowledge in:

  1. Cloning, mutagenesis and other DNA manipulations. 
  2. Become familiar with different recombinant protein expression systems, including troubleshooting.
  3. Introduction to different protein purification techniques
  4. Protein biophysical characterization
  5. Protein crystallization & 3D structure analysis
  6. Protein bioinformatics
  7. Basic understanding of methods to analyse protein-protein interactions

Reading List

N/A

Website

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