Course Identification

New strategies for cloning and protein expression
20243651

Lecturers and Teaching Assistants

Dr. Tamar Unger, Dr. Yoav Peleg, Ms. Meital Yona, Shiran Amir, Ms. Hedva Hamawi, Ms. Eden Levi
N/A

Course Schedule and Location

2024
First Semester
January 28-30, from 13:00-16:30, theory part February 11-15, 18-19 from 13:00-16:30, Hands-on part,
28/01/2024
15/02/2024
43

Field of Study, Course Type and Credit Points

Life Sciences: Laboratory; Elective; Regular; 1.00 points
Chemical Sciences: Laboratory; Elective; Regular; 1.00 points
Life Sciences (ExCLS Track): Laboratory; Elective; Regular; 1.00 points

Comments

This course will be held in person only.

The course's new dates are:
January 28-30, from 13:00-16:30, theory part February 11-15, 18-19 from 13:00-16:30, Hands-on part

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. 

Restrictions

20

Language of Instruction

English

Registration by

22/11/2023

Attendance and participation

Obligatory

Grade Type

Numerical (out of 100)

Grade Breakdown (in %)

10%
90%

Evaluation Type

Examination

Scheduled date 1

10/03/2024
WSoS, Rm C
0900-1200
N/A

Scheduled date 2

26/03/2024
WSoS, Rm C
0900-1200
N/A

Estimated Weekly Independent Workload (in hours)

5

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:

  1. Demonstrate theoretical and practical knowledge in different DNA cloning and mutagenesis methodologies
  2. Conduct experiments using different recombinant protein expression systems, including troubleshooting.

Reading List

DNA manipulations:

  1. 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.
  2. 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.
  3. Unger T and Peleg Y. (2012), Methods Mol Biol 800, 187-199; Recombinant protein expression in baculovirus-infected insect cell system.
  4. Peleg Y and Unger T (2012), Methods Mol Biol 800, 172-186; Resolving bottlenecks for recombinant protein expression in E. coli.

Website

N/A