Gene Cloning Flashcards
What are advantages and disadvantages of bacterial expression system
Advantages-
Fast breeding
Low cost
High expression
Easy to purify
Disadvantages-
Limited ability for post translational modification
Sophisticated protein products may have low biological activity
What are the advantages and disadvantages of yeast expression systems
Advantages-
Need simple media
Can express secretory and intracellular proteins
Disadvantages-
Offers both N and O linked oligosaccharides on proteins (only O in mammals)
What are the advantages and disadvantages for insect expression systems
Advantages-
Can be used in static or suspension culture
Can add post translational modifications
Disadvantages-
Endotoxin accumulation
Protease contamination from host protein leading to degradation
More demanding conditions
What are the advantages and disadvantages of mammalian expression systems
Advantages-
Can express complex proteins
Proteins can be produced transiently by stable expression
Disadvantages-
Demanding cultural conditions
High cost
What are the advantages and disadvantages of algal expression systems
Advantages-
Low costs
High yield due to rapid growth rates
Disadvantages-
Cell walls can be challenging to disrupt making it hard to extract proteins
Technological challenges in optimising the production of specific compounds
What are the advantages and disadvantages of cell free expression systems
Advantages-
Open system so can add unnatural components
Fast expression
Simple
Disadvantages
Limited lifespan due to enzyme stability, resource depletion, metabolic arrest
Technically challenging and complex
What is the central dogma of molecular biology and limits of disipline
The flow of genetic information within a biological systems (DNA MAKES RNA MAKES PROTEIN)
Problems is this is an oversimplification as it’s neglects the complexities of other cellular components/processes (e.g. non coding RNA or post translational mods)
What is the phage group and what was it’s impact on molecular biology
Phage group-
Network of scientists in mid 20th century studying bacteriophages (viruses that infect and replicate in bacteria)
Discoveries-
DNA is hereditary material
That the genetic code is triplet code
Genes encode proteins
DNA is semi conservative
What is protein purification
Isolating protein of interest from a mixture
Steps-
Cell lysis
Separation through chromatography (based on affinity or ion exchange or size)
What is protein crystallography
Technique to determine the 3 dimensional atomic structure of proteins
Proteins can form crystals which can diffract x-rays (the specific diffraction pattern provides info on the spatial arrangement of atoms within the protein)
