New strategies for cloning and protein expression
Lecturers and Teaching Assistants
Dr. Yoav Peleg, Dr. Tamar Unger
Course Schedule and Location
March 19-26, 2023; between 9:30-16:30., FGS, Lab
Field of Study, Course Type and Credit Points
Life Sciences: Laboratory; Elective; Regular; 1.00 points
Chemical Sciences: Laboratory; Elective; Regular; 1.00 points
March 19-26, 2023
The course is aimed to students in Life-Science s and Chemistry. The final format of the course will be determined near the beginning of the course based on the Feinberg policy and regulations. Each student is required to give a 10-15 minutes presentation describing his/her research with an emphasis on topics related to the lab course. The course will be held daily between 9:30-16:30.
Basic knowledge in molecular biology
Attendance and participation
Estimated Weekly Independent Workload (in hours)
The course is intended to provide an overview on various aspects of molecular DNA manipulations and recombinant protein expression. The course covers basic and advanced methods in DNA cloning, mutagenesis and protein expression. Students will have hands-on training on the different steps of cloning and protein expression. Students will be able to work on their own target genes upon prior coordination.
The following topics will be covered:
- Molecular DNA manipulations; cloning and mutagenesis: Cloning using RF (Restriction Free), Transfer-PCR (TPCR), Gibson assembly and Golden Gate assembly.
- Protein expression in E. coli, yeast, baculovirus-infected insect cells and mammalian systems.
- Small scale protein purification.
Upon successful completion of this course students should be be able to:
- Demonstrate theoretical and practical knowledge in different DNA cloning and mutagenesis methodologies
- Conduct experiments using different recombinant protein expression systems, including troubleshooting.
- Unger T, Jacobovitch Y, Dantes A, Bernheim R, Peleg Y (2010), JSB 172, 34-44; Applications of the Restriction Free (RF) cloning procedure for molecular manipulations and protein expression.
- Erijman A, Dantes A, Bernheim R, Shifman JM, Peleg Y. (2011), JSB 175, 171-177; Transfer-PCR (TPCR): a highway for DNA cloning and protein engineering.
- Unger T and Peleg Y. (2012), Methods Mol Biol 800, 187-199; Recombinant protein expression in baculovirus-infected insect cell system.
- Peleg Y and Unger T (2012), Methods Mol Biol 800, 172-186; Resolving bottlenecks for recombinant protein expression in E. coli.