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Genetic Manipulation

Let’s talk about what is DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost
all other organisms.

So now let’s learn what is genetic manipulation, it is the process of inducing changes in gene expression
and expression of novel genes and has proven to be an indispensable tool in genetic research.

According to Bhagvanji, Gohil Sanjay (2018) one of the applications of genetic manipulation is Gene
cloning, defined as a mechanism by which each time the host cell reproduces; several copies of a
particular gene are produced wherein it is possible to clone whole species. Gene Cloning is the insertion
of a fragment of DNA carrying a gene into a cloning vector and subsequent propagation of recombinant
DNA molecules into many copies is known as gene cloning. It involves using bacteria to make multiple
copies of a gene, foreign DNA is inserted into a plasmid, and the recombinant plasmid is inserted into a
bacterial cell, reproduction in the bacterial cell results in cloning of the plasmid including the foreign
DNA and this results in the production of multiple copies of a single gene.

Nuclear cloning or Nuclear transfer- the introduction of the nucleus from a cell into an enucleated egg
cell (an egg cell that has had its own nucleus removed). This can be accomplished through fusion of the
cell to the egg or through the direct removal of the nucleus from the cell and the subsequent
transplantation of that nucleus into the enucleated egg cell. The donor nucleus used for nuclear transfer
may come from an undifferentiated embryonic cell or from a differentiated adult cell (somatic cell); in
the latter case, the technique is called somatic cell nuclear transfer. The concept of nuclear transfer was
first conceived in 1928 by German embryologist Hans Spemann, who initially experimented with
transferring salamander embryonic cell nuclei into egg cells (Rogers, Kara).

Transgenic organism- Modern genetic technology can be used to modify the genomes of living
organisms. This process is also known as “genetic engineering.” Genes of one species can be modified,
or genes can be transplanted from one species to another. Genetic engineering is made possible by
recombinant DNA technology. Organisms that have altered genomes are known as transgenic. Most
transgenic organisms are generated in the laboratory for research purposes. For example, “knock-out”
mice are transgenic mice that have a particular gene of interest disabled. By studying the effects of the
missing gene, researchers can better understand the normal function of the gene (Transgenic
Organisms. 2019. Genetics Generation).

Transgenic organisms have also been developed for commercial purposes. Perhaps the most famous
examples are food crops like soy and corn that have been genetically modified for pest and herbicide
resistance. These crops are widely known as “GMOs” (genetically modified organisms).

Here are few other examples of transgenic organisms with commercial value:

Golden rice: modified rice that produces beta-carotene, the precursor to vitamin A. Vitamin A deficiency
is a public health problem for millions of people around the world, particularly in Africa and Southeast
Asia. Golden rice is still waiting regulatory approval.
Goats that produce important proteins in their milk: goats modified to produce FDA-approved human
antithrombin (ATryn), which is used to treat a rare blood clotting disorder in humans. Goats have also
been genetically modified to produce spider silk, one of the strongest materials known to man, in their
milk. Proposed uses for this recombinant spider silk range from artificial tendons to bulletproof vest.

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The process of cloning a gene in a bacterial plasmid can be divided into five steps:

1. Isolation of vector and gene-source DNA. The source DNA may come from human tissue cells. The
source of the plasmid is typically E. coli. This plasmid carries useful genes, such as ampR, conferring
resistance to the antibiotic ampicillin.

2. Insertion of DNA into the vector. By digesting both the plasmid and human DNA with the same
restriction enzyme we can create thousands of human DNA fragments, one fragment with the gene that
we want, and with compatible sticky ends on bacterial plasmids. After mixing, the human fragments and
cut plasmids form complementary pairs that are then joined by DNA ligase. This creates a mixture of
recombinant DNA molecules.

3. Introduction of the cloning vector into cells. Bacterial cells take up the recombinant plasmids by the
transformation. This creates a diverse pool of bacteria, some bacteria that have taken up the desired
recombinant plasmid DNA, other bacteria that have taken up other DNA, both recombinant and
nonrecombinant.

4. Cloning of cells (and foreign genes).  We can plate out the transformed bacteria on a solid nutrient
medium containing ampicillin.  Only bacteria that have the ampicillin-resistance plasmid will grow.

5. Identifying cell clones with the right gene.  In the final step, we will sort through the thousands of
bacterial colonies with foreign DNA to find those containing our gene of interest

Transgenic animals are used to study diseases and gene functions. Transgenic mice were used to study
development and disease; the first mouse used was called an oncomouse used to study cancer. Other
mice are used to study diabetes, brain function, and development and sex determination. Gene
knockout mice used to study gene function – by purposely “turning off” specific genes. The knockout
mouse does not have a functional gene for a protein called leptin, which helps to control food intake.
Researchers are using this type of mouse to study obesity.