Name: LOCSON, Jan Nheille D.

Section: Cystine A May 23, 2021

MOLECULAR BIOLOGY MIDTERM OUTPUT

Restriction Enzymes

1. Explain what a restriction enzyme is and what it does.

Restriction enzymes, also referred to as restriction endonucleases (REs), are enzymes

isolated from bacteria and other prokaryotes and their isolation was crucial to the development
of recombinant DNA technology (gene cloning or gene splicing) and genetic engineering. REs
recognize and bind to a specific DNA sequence, which called as the restriction site. When
REs find their target DNA sequence, they will create a double-stranded cut in the DNA and
the cut is at or near the restriction site that occurs in an orderly, predictable pattern. Most
bacteria use REs as a defense against bacteriophages, since REs prevent the replication of
the phage by cleaving its DNA at specific sites.

2. Describe what a restriction fragment is and how they are formed.

Restriction fragments of DNA are generated by specific bacterial endonucleases that cut
the DNA though highly defined sequences, which are 4 to 6 bases in length.

For example, EcoRI, a restriction endonuclease isolated from Escherichia coli, recognizes
the following DNA sequence:

5’ GAATTC 3’
3’ CTTAAG 5’

EcoRI cuts the DNA between the G and A on each strand, generating single-stranded
DNA overhangs, which are a stretch of unpaired nucleotides in the end of a DNA molecule.

5’ ---------G 3’ and 5’ AATTC--------- 3’

3’ ---------CTTAA 5’ 3’ G--------- 5’

The fragments created by EcoRI are called as or sticky, since they are complementary at
their single-stranded overhangs and can stick together. Sticky ends are beneficial in gene
cloning because they hold two fragments of DNA together and can be linked by DNA ligase.

3. Give 2 reasons why restriction fragments are important in gene technology.

Restriction fragments are significant in gene technology because they are easier to study,
identify and characterize genes. They are also used in recombining, or joining, DNA molecules
from different genomes, usually with the goal of identifying and characterizing a gene or
studying gene expression and regulation, which provides molecular biologists powerful tools
to create nearly limitless combinations of recombinant DNA.