GEN ERAL

BIOLOGY 2
HSBIO
002

2022-2023
–
PRE-ACTIVITY

Check your understanding

-Jovert Bognot
WHAT IS BIOTECHNOLOGY?
 Technology based on biology
The use of theories and concepts in Biological Sciences to improve our quality of
life.
Biotechnology is simply applied biology. The areas where Biotechnology is mostly used
are in the fields of:
• Medicine
• Agriculture
• Biofuels
• Genetics
What is Genetic Engineering?
• Also called as Recombination, the products are called Recombinant
DNA
• The goal of genetic engineering is to identify traits that are favorable
and insert it in another organism. In this case, we can have animals or
plants with desirable traits or that they can produce specific protein.
• The most common gene editing tool is CRISPR (clustered regularly
interspaced short palindromic repeats)
B T Plants
• One good example of a product of Genetic Engineering is the
production of BT Plants.
• BT stands for the bacteria Bacillus thuringiensis. This is a type of
bacteria that lives in the soil. These bacteria are producing proteins
that are harmful to insects especially several species of Moths and
Butterflies.
• What happens is that the gene responsible for producing these
proteins is introduced to several species of plants and becomes
naturally insect-repellant.
TARGET DNA
• The gene of interest.
• The cell or organism where the target DNA can be found is called a
donor
RESTRICTION ENZYMES
• Used to cut DNA into fragments
• Restriction enzymes are based from bacteria that are immune to
bacteriophages. They cut specific segments of the DNA called the
restriction sites.
• Restriction sites are naturally, 4-8 bases which are palindromic in
nature
• Restriction enzymes may leave sticky ends or blunt ends.
DNA CLONING VECTORS
• carry target gene into a host cell,in example, Plasmids and
Bacteriophage
• Plasmid is circular double stranded DNA in a bacteria. It is
different from the chromosome. Though, plasmids carry-out
DNA molecules, they are not capable of manifesting the trait nor
produce the protein
HOST CELL
• a bacterial cell that allows the
cloning vector to replicate within
it.
• The host should be non-
pathogenic, harmless micro-
organism which is easy for
cultivation
 introduces minor bases
into DNA or RNA e.g, DNA
MODIFYING Ligase and Taq
Polymerase.

ENZYMES Taq polymerase replaces

DNA Polymerase in PCR
METHODS OF GENE
CLONING
 DNA cloning is a molecular biology technique that makes many
identical copies of a piece of DNA, such as a gene. In a

typical cloning experiment, a target gene is inserted into a

circular piece of DNA called a plasmid.

CLONING There are two types of DNA Cloning: (A) Molecular Cloning and
(B) Reproductive Cloning. Dolly the sheep is a by product of a
Reproductive Cloning called SCNT or Somatic Cell Nuclear
Transfer.

Most recent were Huahua and Zhong Zhong, macaques from

china
DNA EXTRACTION
• Before you start with
cloning, you have to
extract first the DNA
from a cell
Stage 1: Isolation of a target gene

• This process can be done using (1)cutting the gene from a complete
chromosome using a restriction enzyme or (2) producing a
complementary DNA (cDNA).
• To do this the DNA must first be cut into fragments and the one containing
the desired gene must be identified.
• Remember to use the same restriction enzyme
Stage 2: Insertion of a t a r g e t
gene into a vector
• Gene is inserted into vectors such as plasmids and
bacteriophages resulting in Recombinant DNA. Bacterial
plasmids that have been isolated from bacterial cells must be
mixed with the same restriction enzymes used to cut the DNA
molecule. This is done to produce the same sequence of sticky
ends.
Stage 3: Introduction of vector
into a host
• The plasmids carrying the target gene must be introduced into a
host cell through transformation, and only a lesser amount will
contain the recombinant plasmid.
Stage 4: Amplification of the t a r g e t
gene by the host c e l l (cloning) ad
screening
• Following the introduction of the recombinant plasmids,
bacteria will then be cultured in a medium. The transformed
bacteria will be able to grow and form colonies on the medium.
The bacteria will divide to produce new identical bacterial cells.
Each time the bacterial cells divide, the recombinant plasmids
will produce multiple copies of the target gene.
POLYMERASE CHAIN REACTION
is a method used to amplify DNA, it resembles
replication but is carried out in a test tube,

TOOLS USED IN PCR

1. Thermocycler 4. primers
2. Test tube containing the DNA sample 5, free DNA nucleotides
3. DNA polymerase (Taq polymerase)
THERMAL CYCLER
• is a laboratory
instrument that heats and
cools samples in
repetitive cycles to
facilitate DNA or RNA
amplification through the
polymerase chain
reaction.
PCR PROCESS
STEP 1: DENATURATION
in which double-stranded DNA templates are heated to
separate the strands;
The reaction temperature is increased to 95 °C, which melts
(disrupts the hydrogen bonds between complementary bases) all
dsDNA into single-stranded DNA (ssDNA).
PCR PROCESS
STEP 2: ANNEALING
The DNA molecule will be exposed at a lower temperature to allow
the primers to attach to the Single-Strand DNA. The role of the primer
is to let Taq Polymerase copy the segment
Taq Polymerase is an enzyme derived from a bacteria Thermus
aquaticus which mimics the job of DNA Polymerase but can
withstand a high temperature
PCR PROCESS
STAGE 3 – EXTENSION
The temperature will be raised again Taq Polymerase will
copy and texted the segment of DNA.

after the third step, it is considered that 1 cycle is complete and

there is 22 amount of DNA in the container (usually an Eppendorf). so
after 1 cycle we can assume that there is 4 copies of DNA.
WHO IS YOUR DESIGNER?