Intro genetic engineering - lecture 12 Flashcards Preview

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What are restriction enzymes?

Cut phosphodiester bonds in DNA at specific sequences


Why do bacteria produce restriction enzymes?

To combat phage infection


EcoRv nuclease vs EcoRV methylase

EcoRv methylate is an enzyme which adds methyl groups to specific sequences , which prevents EcoRv nuclease form cutting there


DNA ligase

catalyse the formation of phospodiester bonds between a 3' hydroxyl and a 5' phosphate group


What are DNA vectors?

Units that can be used to store, replicate and manipulate genetic info . Plasmids are commonly used.


What are expression vectors

Have a promoter near a multiple cloning site


How do you get a gene of interest into a plasmid/

- Restriction enzymes at end of gene
- Design PCR primer to have restriction enzymes at the end


what is the purpose of site directed mutagenesis?

Used to introduce specific changes within. the DNA sequence


How does site - directed mutagenesis occur?

- methylated template plasmid to start with
- Do PCR and you get a plasmid with no methylation
- Dnp1 restriction enzyme degrades methylated temple plasmid
- left with vector with mutation


The Cre- Loxp system

- Use of the Cyclization recombinase (cre) enzyme produced by bacteriophages
- Cre recognises Loxp sequences and induces a deleted sequence then recombination


What is Crispr - Casp?

- A cellular systems which exists in bacteria to combat viruses
- components have been humanised to edit human genomes


What does Palindromic mean?

n genetics, a DNA or RNA sequence that reads the same in both directions. The sites of many restriction enzymes that cut (restrict) DNA are palindromes.


What organism uses the Cre-Lox P system?