Recombinant Protein Expression
Run by School of Natural Sciences
10.000 Credits or 5.000 ECTS Credits
Organiser: Dr Olga Golyshina
Overall aims and purpose
One of the most important aspects of Biotechnology is the production of recombinant proteins. We will address this topic in this module by examining strategies for the production of recombinant proteins in bacteria – these will include plasmid expression vectors, the choice of promoters designed to drive expression, cloning suitable DNA sequences into these plasmid vectors, transformation and selection of recombinant bacteria, and the analysis of recombinant proteins on polyacrylamide gels. The theory of recombinant protein production will be covered in a number of lectures which are linked to the practicals.
1)This module will address aspects of the production of recombinant proteins in bacteria. 2)These will include details of plasmid vectors used to drive expression in different hosts. 3) Details of promoters and leader sequences which can be used to both drive expression and details of secretory pathways in prokaryotic and eukaryotic cell systems. 4) Cloning and selection strategies to insert suitable coding sequences downstream of suitable promoters in plasmid vectors will be covered, and the analysis of recombinant plasmids will be detailed. 5)Cloning, purification and analysis of the recombinant proteins produced in bacterial systems will be covered in the practical sessions allocated to this module. 6) Lecture material will, in addition, use the topic of microbial diversity to study specific examples of the production of biologically important recombinant proteins.
Written reports and presentations should demonstrate at least a basic knowledge and understanding of the essential facts and key concepts of all the subjects covered in this module
Complete familiarity with the subject, showing confidence in handling pertinent facts and arguments. Demonstration of assimilation and understanding of the material gleaned from a variety of sources including lectures, books, reviews and articles in the scientific literature. Ability to present material with a strong element of originality, individuality and insight and to integrate it into a broad biological context. Conveys a convincing impression of being an authority on the subject.
Thoroughly competent appreciation of the subject with no major errors or misunderstandings. Presentation of the material in a logical and ordered manner. The report would usually be expected to show evidence of gleaning information from background reading, although an excellent synthesis of the lecture material could gain marks towards the lower end of this category. No demonstration of great originality.
Knowledge of the methods of isolating and analysing recombinant proteins on polyacrylamide gels.
Knowledge and understanding of bacterial transformation.
Knowledge and understanding of the selection of recombinants and the characterisation of recombinant plasmids.
Critical understanding of the cloning strategies used to create a recombinant expression plasmid.
A knowledge of different promoters which can be used to alter expression levels.
Detailed knowledge of the plasmid vectors used to express recombinant proteins.
|Presentation on heterologous expression hosts||50.00|
Teaching and Learning Strategy
5 X 1 hour lectures, twice a week.
5 x3 hour practicals, to run sequentially.
Students will also be directed to the relevant primary literature, book sections and web resources, which they will be expected to read during the allotted private study time.
- Literacy - Proficiency in reading and writing through a variety of media
- Numeracy - Proficiency in using numbers at appropriate levels of accuracy
- Computer Literacy - Proficiency in using a varied range of computer software
- Self-Management - Able to work unsupervised in an efficient, punctual and structured manner. To examine the outcomes of tasks and events, and judge levels of quality and importance
- Exploring - Able to investigate, research and consider alternatives
- Information retrieval - Able to access different and multiple sources of information
- Inter-personal - Able to question, actively listen, examine given answers and interact sensitevely with others
- Critical analysis & Problem Solving - Able to deconstruct and analyse problems or complex situations. To find solutions to problems through analyses and exploration of all possibilities using appropriate methods, rescources and creativity.
- Safety-Consciousness - Having an awareness of your immediate environment, and confidence in adhering to health and safety regulations
- Presentation - Able to clearly present information and explanations to an audience. Through the written or oral mode of communication accurately and concisely.
- Argument - Able to put forward, debate and justify an opinion or a course of action, with an individual or in a wider group setting
- Self-awareness & Reflectivity - Having an awareness of your own strengths, weaknesses, aims and objectives. Able to regularly review, evaluate and reflect upon the performance of yourself and others
Subject specific skills
- Engagement with current developments in the biosciences and their application.
- Appreciation of the complexity and diversity of life processes through the study of organisms.
- Engage in debate and/or discussion with specialists and non-specialists using appropriate language.
- Undertake field and/or laboratory studies of living systems.
- Undertake practical work to ensure competence in basic experimental skills.
- Demonstrate awareness of the importance of risk assessment and relevant legislation
Resource implications for students
Courses including this module
Compulsory in courses:
- C7AA: MSc Molecular Biology with Biotechnology year 1 (MSC/MBB)