Biotechnology the DNA exponentially for 25 to 75

cycles.
Tools used in Genetic Engineering
 A cycle takes only a minute, and each
 Genetic engineering involves new segment of DNA that is made can
manipulating genetic material (DNA) serve as a template for new ones. This
to achieve the desired goal in a technique is used in molecular biology
predetermined way. to amplify a single copy or a few copies
 Genetic engineering also made cloning of a segment of DNA across several
possible as it successfully cloned a orders of magnitude, generating
mammal from an embryo cell, a sheep thousands to millions of copies of a
named “Dolly,” and Ian Wilmut and his particular DNA sequence.
colleagues executed the said cloning.  Developed in 1983 by Kary Mullis, PCR
is now a common technique used in
Genetic Engineering Defined clinical and research laboratories for
 The term genetic Engineering is initially various applications (Javed, 2017).
referred to various techniques used for Restriction Enzymes (Molecular Scissor)
the modification or alteration of
organisms through the processes of  are enzymes that create one
heredity and reproduction. incision on each of the two strands
of DNA at specific locations based
Genetic Engineering In Agriculture on the nucleotide sequence.
 Can use genetic engineering to  DNA cut with a restriction enzyme
introduce a new sequence of DNA into produces many smaller fragments
plants making them more resistant to of varying sizes.
insects Bacillus Thuringiensis (BT) crops.  These can be separated using gel
 To make crops more resistant to pests. electrophoresis or chromatography.
 Genetically engineered crops that  Restriction Enzymes was isolated in
contain bacterium crystal toxins that 1970 by Hindll. He also did the
mak4 them more resistant to other subsequent discovery and
insects. characterization of numerous
restriction endonucleases (Hitendra,
Human Insulin 2018)
 Medicine for diabetes Gel Electrophoresis
 Compost of E.coli and yeast
 Thansgenic or genetically modifies  s is used for various purposes, from
viewing cut DNA to detecting DNA
Genetically Modified Organisms (GMO) inserts and knockouts. It is also used to
 An organism generated through genetic estimate the molecular weight of
engineering. protein and nucleic acids, purification of
isolated proteins, monitoring changes of
THE DIFFERENT TOOLS USED IN GENETIC protein content in body fluids, blotting
ENGINEERING application, and many more.
 Purifying DNA from cell culture or
 Polymerase Chain Reaction (PCR) is
cutting it using restriction enzymes
efficient technique because it multiplies
 would not be of much use if we could genes that results from the
not visualize the DNA that is, find a way multiplication of a single cell,
to view whether or not your extract organism, or gene.
contains anything or what size  Has no thru nucleus
fragments you have cut it into. One way
Eukaryostic Host
to do this is by gel electrophoresis
(Tapeshwar, 2015).  Produce human proteins
(human insulin)
DNA Ligase
 They have thru nucleus
 are enzymes that can create  The most commonly used
covalent bonds between nucleotide eukaryotic organism is the
chains. yeast, Saccharomyces
 Link two or more individual strands cerevisiae. It is a non-
for DNA to create a recombinant pathogenic organism
strand or close to a circular strand. routinely used in the
brewing and baking
Polymerase
industry. Certain fungi have
 The group of enzymes that also been used in gene
catalyze the synthesis of nucleic cloning experiments (Faraza
acid molecules ,2017).

Types of Polymerase Selection of Small Self-Replicating DNA

 DNA-dependent DNA  Small circular pieces of DNA that are not

polymerase that replicates DNA part of a bacterial genome, but are
from DNA capable of self-replication, are known as
plasmids
 RNA-dependent DNA  Plasmids are often used as vectors to
polymerase (reverse transport genes between
transcriptase) that transcribes microorganisms
DNA from RNA.  Viral (bacteriophage) DNA can also be
used as a vector, as can cosmids,
 DNA-dependent RNA recombinant plasmids containing
polymerase that transcribes bacteriophage genes (Faraza ,2017)
RNA from DNA
Transformation

 Process of transferring genetic

Prokaryotic Host
material ona vector (such as
 are able to multiply their plasmid) into new cells.
plasmids (along with foreign  This technique requires the host
DNA) also multiply to produce cells to be exposed to an
millions of copies, referred to as environmental change making them
a colony or in a short clone. “competent” or temporarily
 The term ‘clone’ broadly refers permeable to the vector.
to a mass of cells, organisms, or
 Electroporation

 The larger the plasmid, the

lower the efficiency which is
taken up by the cells.

Transduction

 Larger DNA segments are move

easily cloned using
bacteriophage, retrovirus, or
other viral vectors.
 Phage or visual vectors are
often used in regenerative
medicine but may cause the
insertion of DNA in parts of our
chromosomes.

Methods to Select Transgenic Organisms

 According to Faraza (2017), not all

cells will take up DNA during
transformation.

ORGANISMS

 Plasmids carry genes for antibiotic

(fight infection) resistance
 Transgenic cells can be selected
based on the expression of these
genes and their ability to grow on
media containing antibiotics.
 Green fluorescence protein allow
the selection based on fluorescence
(emit or give of light).