Enzyme and restriction mapping Flashcards
What type of proteins is genetic engineering use to produce?
- Recombinant proteins
- Insulun
- Interferon
- G-CSF - Transgenic organisms
- Disease models
- Improved agricultural yields
What do nucleases degrade and how?
Degrade nucleic acids by hydrolysing phosphodiester bonds
What does ribonuclease degrade?
Degrades RNA
What does DNase degrade?
Degrades DNA
What does exonuclease degrade?
Degrade from the end of the molecule
What does endonucleases degrade?
Degrade within nucleotide chain
What are the 2 things that restriction endonucleases do?
- Recognise a specific sequence
2. Cut that sequence
Physiology behind restriction
Limit transfer of nucleic acids from infecting phages into bacteria
Where are the many different enzymes of restriction endonucleases from?
Many different enzymes from different bacteria
What can some nucleases produce?
Some nucleases produce an overhang and some produce a blunt end
What are restriction maps?
Map of restriction sites within a molecule
What way is useful for describing plasmids?
Restriction maps are a useful way of describing plasmids
Restriction enzymes in molecular diagnostics
Sickle cell anaemia:
- Single point mutation
- A–>T resulting in Glu–>Val
What can single nucleotide changes do?
Single nucleotide changes can create/destroy restriction enzyme sites
What does DNA ligase do and how?
-Puts 2 fragments together
Does this by repairing nicks in phosphodiesterase backbone
