Restriction Enzymes

enzymes

are biological catalysts that are found in all cells

enzymes

they are molecules that speed up chemical reactio

enzymes

have unique chemical structures that mean they only act on specific substrates

temperature, pH

for a particular enzyme to work optimally, it must be under specific conditions (— —) otherwise its unique chemical structure will be disrupted

enzymes

are critical for a range of cellular processes including digestion, DNA replication, protein synthesis

1. making recombinant DNA and appraising success

2. DNA profile analysis

restriction enzymes purposes

Bacteria

The origin of enzymes

Restriction enzymes

Used to help bacteria destroy viral DNA

Gel electrophoresis

Used to seperate pieces of DNA for analysis

Restriction enzymes

It cuts between specific bases (letters) of the doulbe stranded DNA molecule. DNA is then in multiple pieces

Restriction endonucleases

They cut within the molecule, another term for restriction enzymes.

Restriction endonucleases

The tool used to cut DNA into small fragments before sequencing it.

Restriction enzymes sites

Specific restriction enzymes cut at specific DNA sequences.

EcoRI

Enzyme that cuts at the sequence :GAATTC

E.Coli

The origin of EcoRI

Restriction fragments

The resulting pieces of DNA after being cut at specific DNA sequences

yes

study slide 11 (derivation of the EcoRI name) type yes

Influenza

Origin of HindIII

P stuartii

PstI

E.coli

Origin of EcoRV

HindiIII

Has 5 protruding ends

PstI

has 3 protruding ends

EcoRV

has blunt ends

1. Used to manipulate DNA for different scientific applications
   2. Used to assist insertion of genes
   3. Used to distinguish gene alleles

Applications of restrictions enzymes

Single nucleotide polymorphisms

Single base changes in DNA

Pseudomonoas syringae

ice-minus bacteria prevent frost crystals from forming in plants

Pseudomonas fluorescens

Nonpathogenic bacterium that can produce proteins rapidly. Advantageous in developing biotherapeutics and vaccines

Agrobacterium tumefaciens

Tumor inducing plasmid that causes crown gall disease in plants. Produces beneficial genes to plants