Course Identification

New strategies for cloning and protein expression
20173182

Lecturers and Teaching Assistants

Dr. Tamar Unger, Dr. Yoav Peleg
N/A

Course Schedule and Location

2017
Second Semester
15.03.2017-23.3.17, 9:30-16:30, FGS, Lab
15/03/2017
23/03/2017
50

Field of Study, Course Type and Credit Points

Life Sciences: Laboratory; Elective; 0.50 points

Comments

The course is aimed to students [prederably PhD students] in Life-Sciences and Chemistry. 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

20

Language of Instruction

English

Attendance and participation

Obligatory

Grade Type

Pass / Fail

Grade Breakdown (in %)

70%
30%

Evaluation Type

Laboratory

Scheduled date 1

N/A
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-
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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 using RF (Restriction Free) and Transfer-PCR (TPCR), Simultaneous insertion & deletion, Multiple-site mutagenesis.
  • Protein expression in E. coli, yeast, baculovirus-infected insect cells and mammalian systems.

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

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