Intro genetic engineering - lecture 12 Flashcards Preview

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Flashcards in Intro genetic engineering - lecture 12 Deck (13)
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1

What are restriction enzymes?

Cut phosphodiester bonds in DNA at specific sequences

2

Why do bacteria produce restriction enzymes?

To combat phage infection

3

EcoRv nuclease vs EcoRV methylase

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

4

DNA ligase

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

5

What are DNA vectors?

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

6

What are expression vectors

Have a promoter near a multiple cloning site

7

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

8

what is the purpose of site directed mutagenesis?

Used to introduce specific changes within. the DNA sequence

9

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

10

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

11

What is Crispr - Casp?

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

12

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.

13

What organism uses the Cre-Lox P system?

bacteriophages