GENERAL

BIOLOGY 2
ALYZA B. DURAN
Teacher 1
01 Enumerate the steps of Recombinant DNA
Technology

02 Outline the process of genetic engineering

techniques

Explain how recombinant organisms can

03 affect a population or an ecosystem
Spot the normal mouse and the genetically
modified mouse
 Methods to modify genes

Insert new Replacement of Removal of Mutation of

genes genes genes existing
(recombination) genes
 7
1
2 3 4 5 6
 Sequencing the gene
Isolating
of DNA Amplifying the
7
Selection of an
recombinant DNA
1
appropriate vector

2 3 4 5 6

Ligation of gene of Selection process to

Cutting or Cleavage screen which cells
of DNA by Restriction interest with the
vector contain the gene of
enzymes interest
 ACTIVITY
DIRECTIONS: Analyze the Diagram. Using the
word choices provided in the boxes, fill in the
numbered boxes with the steps of bacteria
transformation and the lettered lines with the
name of the structure next to it.
Using the word choices provided in the boxes, fill in the numbered boxes with the steps of
bacteria transformation and the lettered lines with the name of the structure next to it.
RECOMBINANT DNA
The newly generated DNA molecules from two different
species that are inserted into a host organism
Why are
plasmids from
bacteria are
commonly used
as vectors?
Why are plasmids from bacteria are
commonly used as vectors?
• carry genes that provide the host cell with useful properties,
such as drug resistance, mating ability, and toxin production.
1

• small enough to be conveniently manipulated experimentally

2

• carry extra DNA that is spliced into them

3
 RECOMBINANT DNA TECHNOLOGY
It refers to the joining
together of DNA
molecules from two
different species that
are inserted into a
host organism to
produce new genetic
combinations.
 Sequencing the gene
Isolating
of DNA Amplifying the
7
Selection of an
recombinant DNA
1
appropriate vector

2 3 4 5 6

Ligation of gene of Selection process to

Cutting or Cleavage screen which cells
of DNA by Restriction interest with the
vector contain the gene of
enzymes interest
Isolating the DNA
 Cutting Or Cleavage Of DNA by
Restriction Enzymes
Restriction Enzymes
A restriction enzyme is a
protein that recognizes a
specific, short nucleotide
Restriction site sequence and cuts the
Restriction DNA only at that specific
site Restriction site, which is known as
enzymes restriction site or target
sequence.
Cutting Or Cleavage Of DNA by
Restriction Enzymes

Restriction site
Restriction
site Restriction
enzymes
Why do bacteria
have restriction
enzymes?
“overhangs”
 Does all restriction enzymes
produce sticky ends?
“blunt cutters”

Result: harder to ligate

 ● Restriction enzymes are named

Where do
for prokaryote species they are
isolated from.

restriction
enzymes get
these weird
names?
 Selection of an appropriate Vector

● Vector should possess an

origin of replication so that it
can self-replicate inside the
host cell. It should have a
restriction site for the
insertion of the target DNA.
It should have a selectable
marker with an antibiotic
resistance gene that
facilitates screening of the
recombinant organism
Ligation of the gene of interest
How does DNA ligase
do this?
 How does DNA ligase do
this in Recombinant DNA?
 Goal : To use the enzyme EcoR1 to insert the gene into the
plasmid.

Using ATP as an energy source, ligase catalyzes a

reaction in which the phosphate group sticking off the
5’ end of one DNA strand is linked to the hydroxyl
group sticking off the 3’ end of the other.
 Amplifying the Recombinant DNA

● Transfer of the
recombinant plasmid
into a host cell (that
would carry out
replication to make
huge copies of the
recombined plasmid).
 Selection process to screen which
cells contain the gene of interest

Plasmid-containing bacteria colony

 Sequencing of the gene to find out the
primary structure of the protein

determines the
exact sequence of
nucleotides, or
bases, in a DNA
molecule.
Why do we need to check colonies?
Using the word choices provided in the boxes, fill in the numbered boxes with the steps of
bacteria transformation and the lettered lines with the name of the structure next to it.
 Do you think
recombinant organisms
could also pose a threat
to a population or
ecosystem?
 Limitations of Recombinant DNA
Technology

● Destruction of native species in the environment the

genetically modified species are introduced in.
● Resilient plants can theoretically give rise to resilient
weeds which can be difficult to control.
● Recombinant organisms contaminating the natural
environment.
 Limitations of Recombinant DNA
Technology

● Ethical concern about humans trying to play

God and mess with the nature’s way of
selection.
● Such a system might lead to people having their
genetic information stolen and used without
permission.
HOME-BASED LEARNING TASK
Direction: Find any article about the advancement of any
country in Asia particularly the Philippines in the field of Genetic
Engineering. You may find the article in the newspaper, a
magazine, a science journal, websites or any other sources that
highlights biotechnology. Use the format below and attach the
article. Answer the following guide questions.
Title: ________________________________________________
Author: ______________________________________________
Source: ______________________ Date: _______________
GUIDE QUESTIONS:
1. Identify the main idea of the article. Write one or two
sentences that summarize each main idea.
2. Identify and define at least five terms that are either new to
you or are related to biotechnology/genetic engineering.
3.What questions does this article raise? Does the article
present differing viewpoints? If so, compare and contrast each
point of view.
4. What is your response to the article? Do you accept the
findings? Why or why not?