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2019/20 Taught Postgraduate Module Catalogue

BIOL5373M Protein Engineering Laboratory Project

15 creditsClass Size: 80

Module manager: Dr Chi Trinh
Email: C.H.Trinh@leeds.ac.uk

Taught: Semesters 1 & 2 View Timetable

Year running 2019/20

Pre-requisite qualifications

BSc in Biological Sciences or equivalent

Pre-requisites

BIOL5292MBioscience MSc Research Project

Module replaces

BIOL5206M

This module is not approved as an Elective

Objectives

- To provide practical training in a range of modern biological experimental techniques principally based around molecular biology.
- To provide training in maintaining laboratory records and in writing scientific research papers.

Learning outcomes
On completion of this module, students should be able to:
- demonstrate an understanding of and practical competence in a range of experimental techniques including: plasmid DNA preparation and quantification; subcloning; PCR; primer design for site-directed mutagenesis, site directed mutagenesis; DNA sequencing; recombinant protein expression; SDS PAGE and western blotting; protein purification and protein analysis;
- maintain a detailed and accurate laboratory notebook recording the experimental procedures and results obtained during the practical sessions;
- present and critically analyse the results obtained in the form of a short scientific paper.


Syllabus

This module is an extended practical investigation in the form of a mini project. Beginning with the pET23a plasmid containing the open reading frame of the Green Fluorescent Protein (GFP), students will subclone the GFP insert into the pET28c protein expression vector and transform into E. coli DH5a cells. Transformed cells will be selected by plating on LB medium containing antibiotic followed by direct colony PCR to detect the presence of the GFP insert.

Students will then go on to generate 2 spectral variants (mutations) of GFP using site-directed mutagenesis and these will be characterized by DNA-sequencing.

Isolated plasmid DNA derived from each of these recombinants will be transformed into the expression host BL21(DE3) and GFP expression induced using the auto-induction method. Protein expression will be assessed by SDS-polyacrylamide gel electrophoresis followed by western blotting. GFP will be purified using NiTA chromatography and then analysed by mass spectroscopy and spectral analysis.

Students will be expected to record their experimental procedures and results in a laboratory notebook and will be given guidance on how to maintain suitable records.

Teaching methods

Delivery typeNumberLength hoursStudent hours
In Course Assessment11.001.00
Lecture21.002.00
Practical33.009.00
Practical86.5052.00
Tutorial11.001.00
Private study hours85.00
Total Contact hours65.00
Total hours (100hr per 10 credits)150.00

Private study

Students should note that the following information is for guidance only. The actual time required for the various elements will vary between students.
- 2.5hrs preparation per practical: 27.5 hours
- Primer-design activity: 3.5 hours
- Preparation of end of module test: 5 hours
- Preparation of short literature review and research paper: 50 hours

Opportunities for Formative Feedback

Attendance at weekly practical classes and quality of literature review.
Students will also be required to keep a contemporaneous record of procedures and results in the form of a laboratory note book which will be marked during the practical session.

Methods of assessment


Coursework
Assessment typeNotes% of formal assessment
Literature ReviewFormative assessment (800 words literature review)0.00
In-course AssessmentEnd of module test - on material covered in pre-practical questions35.00
In-course AssessmentQuality and completeness of information recorded in laboratory note book15.00
Research ProposalPresentation and critical analysis of results obtained in the form of a short research paper50.00
Total percentage (Assessment Coursework)100.00

Normally resits will be assessed by the same methodology as the first attempt, unless otherwise stated

Reading list

The reading list is available from the Library website

Last updated: 15/08/2019

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