Course Identification
New strategies for cloning and protein expression
Lecturers and Teaching Assistants
Dr. Tamar Unger, Dr. Yoav Peleg, Ms. Meital Yona
Course Schedule and Location
Both the theoretical session and the lectures during the Hands-on session will be held in the teaching room next to the lab.,
23/02/2025
37
Field of Study, Course Type and Credit Points
Life Sciences: Laboratory; Elective; Regular; 1.00 points
Chemical Sciences: Laboratory; 1.00 points
Comments
This course will be held in person only.
23-25.2 - theoretical part (13:00-16:00)
9-12.3 and 16-18.3 - Hands-on part (13:00-17:00).
Both the theoretical session and the lectures during the Hands-on session will be held in the teaching room next to the lab.
Students will work on their own gene targets in addition to the course projects.
The course is aimed to students in Life-Sciences and Chemistry and will cover topics in DNA manipulation and recombinant protein expression in bacteria and human cells. The course is divided into two parts: Theoretical part on cloning/mutagenesis strategies and primer design. Practical part will include hands-on work on individual students projects as well as course projects.
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.
Prerequisites
1) Basic knowledge in molecular biology.
2) Each student needs to have the SnapGene software on its own laptop.
The software can be obtained through the Weizmann IT. This software is obligatory for primer design.
Registration by
01/12/2024
Attendance and participation
Scheduled date 1
18/03/2025
Estimated Weekly Independent Workload (in hours)
Syllabus
The course will expose students to the different methods in cloning/mutagenesis and protein expression. Students will have the opportunity to work on their own target genes and to design the primers, as well as, to express the proteins, if applicable. The course is intended to provide an overview on various aspects of molecular DNA manipulations and recombinant protein expression.
The following topics will be covered:
- Molecular DNA manipulations; cloning and mutagenesis: Cloning using RF/TPCR (Restriction Free/ Transfer-PCR), In-Fusion, Inverse-PCR etc.
- Protein expression in E. coli, baculovirus-infected insect cells and mammalian systems.
- Small scale protein purification.
Learning Outcomes
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.
Reading List
DNA manipulations:
- 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.