Course Identification

Title:

New strategies for cloning and protein expression

Code:

20213332

Lecturers and Teaching Assistants

Lecturers:

Dr. Tamar Unger, Dr. Yoav Peleg, Ms. Meital Yona, Mr. Asa Tirosh, Shiran Amir

TA's:

N/A

Course Schedule and Location

Year:

2021

Semester:

Second Semester

When / Where:

March 11-18; 9:30-16:30, FGS, Lab

First Lecture:

11/03/2021

End date:

18/03/2021

No. of hours (laboratories):

38

Field of Study, Course Type and Credit Points

Life Sciences: Laboratory; Elective; Regular; 0.50 points
Chemical Sciences: Lecture; Elective; Regular; 0.50 points
Life Sciences (Molecular and Cellular Neuroscience Track): Lecture; Elective; Regular; 0.50 points
Life Sciences (Computational and Systems Biology Track): Lecture; Elective; Regular; 0.50 points

Comments

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.

Prerequisites

Basic knowledge in molecular biology

Restrictions

Participants:

20

Language of Instruction

English

Registration by

Registration By:02/02/2021

Attendance and participation

Obligatory

Grade Type

Pass / Fail

Grade Breakdown (in %)

Attendance

70%

Final:

30%

Evaluation Type

Laboratory

Scheduled date 1

Date / due date

N/A

Location

N/A

Time

-

Remarks

N/A

Estimated Weekly Independent Workload (in hours)

5

Syllabus

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.

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.

Website

N/A