Molecular Biotechnology

BGY9003M

Introduction

James Flint: jflint@lincoln.ac.uk

 Module lead : James Flint
• Senior Lecturer in Biotechnology
– Carbohydrate active enzymes
– Industrial application of biotechnology/proteins
– Industrial Liaison for School of Life Sciences

JBL2W12
Ext 6804
jflint@lincoln.ac.uk
Office hours : Tuesday 10:00-12:00
 Blackboard Site
• First port of call
Module announcements
for all module to whole cohort
information Information on lectures
and general module

Moderated peer
Normally!! discussion
 Module Aims

“designed to give a sound basis in the theory

and practical application of the molecular and
bioinformatics techniques heavily used in
modern biotechnology ”
 Teaching Staff
• Module leader
– James Flint
• Teaching staff
– Edward Taylor + Other experts
– Ross Williams – GE Healthcare
– Alan Goddard – Tissue and Organ culture
– Stefan Millson
– Elizabeth Mitchell
– Stephen Prior
 Module Content
• Methods based
• Detailed knowledge of:
– molecular biology,
– structural biology,
– biochemistry,
– bioinformatics,
– cell biology,
– biophysics
 Module Content
• 23 Lectures
– Molecular biology
– Nucleic acid techniques
– Cell culture techniques
– Protein techniques
– Biophysics and functional characterisation
• 12 Practicals
– Wet and dry practicals
 Lecture
Week Specific content
number
1 1 Introduction
2 Nucleic acids, proteins, carbohydrates and lipids
3 Central dogma, pathways and metabolism
2
4 Nucleic acid isolation and visualisation
5 Hybridisation - Southern, Northern, FISH, RNAi
3
6 PCR - varieties and mutagenesis
7 Vectors, cloning and reporters
4
8 Sequencing technologies
9 Yeast 2-hybrid systems
5
10 Fermentation
11 Recombinant protein expression
6
12 Cell lines and immortalisation
Lectures

13 Cell fractionation
7
14 Tissue and organ culture
15 Protein purification
8
16 Visualization- 1D and 2D protein electrophoresis
17 Structure determination -crystallography
9
18 Lipids in Biotechnology
19 Structure determination -NMR and Cryo-EM
10
20 SPR, microcalorimetry, CD
21 Spectroscopy- Fluorescence, Absorbance and Raman
11
22 BLAST, ClustalW, Expasy, EMBI
23 Biochemical assays
16
24
 Outline
Basic skills 1
Basic skills 2
Bacterial gDNA isolation
PCR
Agarose gel electrophoresis
Cloning and transformation
Plasmid isolation, digestion and
Wet
electrophoresis
Practicals

SDS-PAGE
Protein purification
NMR

Outline
Dry

Bioinformatics and primer design

Sequencing and sequence analysis
Practical outline
 Practical style
• Different to undergraduate
– May be given a full protocol with background
– May be given a set of verbal instructions
– May be given SOP
– May be given a paper to base practical off
– May be more like a workshop

• In “real world” all of these are possible

 Assessment
• Assessment 1. Write up of practicals 3-12 as
research article (2500 words max)
– In journal style

Abstract
Introduction
Methods
Results and discussion
Bibliography (not included in word count)
 Help with writing a lab report
• Peer assisted learning
– Working with your colleagues
• Formative Assessment. Write up of practicals
3-6 as research article (1500 words max)
– Style
– Content
– Background
 Assessment
• Assessment 2. Written proposal of strategy
directed to a line manager on how to address
a biotechnologically relevant problem (5000
words max)

LincProt Incorporated want to make a clopyralid

recognition protein for incorporation into a
biosensor device. How should they go about it?
 Remit of sensor assignment
• THERE IS NO “RIGHT” ANSWER
• Should be able to work in aqueous
environment
• Rapid and/or sensitive detection
– Which is more important?
• Needs to detect clopyralid but “could” detect
other things (??)
– Specificity vs sensitivity?

Justify your choices!!

 Potential inclusion
Protein(s) – which
and why? Validation/
characterisation Expression system
– which and why?

Cost/feasibility
Limitations
Risks
Specificity

Novelty Type of Biosensor–

which and why?
Technology
 Deadlines
• Formative Assessment – Lab report
– 16:00 17th November 2017

• Assessment 1 – Lab Report

– 16:00 19th January 2018
• Assessment 2 – PBL strategy document
– 16:00 26th January 2018
Any questions?