BIO-SCOPE SUMAN DEBNATH (M.Sc in ZOOLOGY), B.

Ed (Life Sciences)

BIOTECHNOLOGY AND ITS APPLICATIONS

The applications of biotechnology include therapeutics, diagnostics, processed food, bioremediation,
waste treatment, energy production, Transgenic animals, Genetically modified crops (more tolerant to
diseases, pests and abiotic stresses like cold, heat, drought), etc.
Research areas of biotechnology:
 Providing the best catalyst in the form of improved organism; generally a microbe or pure enzyme.
 Creating optimal conditions through engineering for a catalyst to act.
 Downstream processing technologies to purify the protein/organic compound.
Genetically Modified Organisms (GMOs)
The organisms which contain functional foreign gene experimentally introduced into their genome by
genetic engineering from another_ species are called genetically modified organisms or transgenic
organisms or transgenics. Foreign gene that alters the host cell genetically is termed as transgene and
production of transgenic organism is known as transgenesis.
Transgenic plants
The plants in which foreign gene has been introduced through genetic engineering are called transgenic
plants. Transgenic plants can be obtained by combination of tissue culture and genetic engineering.
 The vector used to introduce new genes into plant cells is most often a plasmid from the soil bacterium
Agrobacterium tumefaciens. This is the Ti plasmid (tumour inducing plasmid), so called because in
nature, it induces tumours in broad leaf plants.

 For using Ti plasmid as a vector, researchers have eliminated its tumour causing properties while
keeping its ability to transfer DNA into plant cells. The part of Ti plasmid transferred into plant cell
DNA, is called the T-DNA.
Production of Pest Resistance Crops:
 Bt cotton is produced by a bacterium called Bacillus thuringiensis (Bt for short). Bt toxin gene has been
cloned from the bacteria and is expressed in plants to provide resistance to insect without the need for
insecticides.
 The Bt toxin protein exists as inactive protoxin but once an insect ingests the inactive toxin, it is
converted into an active form of toxin due to the alkaline pH of the alimentary canal which solubilises
the crystals. The activated toxin binds to the surface of midgut of epithelial cells and creates pores that
cause cell swelling and lysis and eventually cause death of the insect.
 Bt toxins are insect-group specific. The toxin is coded by a gene named cry. Two cry genes (cry I Ac)
and (cry II Ab) have been incorporated in cotton. This genetically modified crop is called Bt cotton as it
contains Bt toxin genes against cotton bollworms.

 Bt brinjal is a transgenic brinjal (also known as an eggplant or aubergine), created by inserting a crystal
protein gene (cry I Ac) from the soil bacterium Bacillus thuringiensis into the genome of various brinjal
varieties.

 Bt corn is a transgenic corn created by inserting cry I Ab to protect the same from corn borer.

Contacts: Suman Debnath, Ph-7687931008. E-Mail: [email protected], Class: XII (NEET) Page 1
BIO-SCOPE SUMAN DEBNATH (M.Sc in ZOOLOGY), B.Ed (Life Sciences)

Protection against nematodes:

A nematode Meloidogyne incognita infects roots of tobacco plants cause great reduction in yield.
RNA interference (RNAi)
 This method involves silencing of a specific mRNA due to a complementary ds RNA molecule that
binds to and prevents translation of the mRNA (silencing).
 The source of this complementary RNA could be from an infection by viruses having RNA genomes or
mobile genetic elements (transposons) that replicate via an RNA intermediate.
 Using Agrobacterium vectors, nematode-specific genes were introduced into the host plant.
 The introduction of DNA was such that it produced both sense and anti-sense RNA in the host cells.
 The anti-sense RNA and mRNA molecules will anneal to form duplex RNA molecules (or double
stranded RNA) and the duplex RNA molecules cannot be translated.
 Thus, silenced the specific mRNA of the nematode.
 The consequence was that the parasite could not survive in a transgenic host expressing specific
interfering RNA.

Production of Improved Varieties

 Golden rice a variety of Oryza sativa rice produced through genetic engineering to biosynthesize
β-carotene, a precursor of pro-vitamin-A in the edible parts of rice. Golden rice was developed as a
fortified food to be used in areas, where there is a shortage of dietary vitamin-A.
 Genetically modified tomato: Flavr Savr an improved tomato variety is developed by the use of
antisense RNA technology. The enzyme polygalacturonase, which damages pectin is deactivated
resulting in increased shelf life.
 Canola oil: is the variety of rapeseed (Brassica napus) crossed with field mustard (Brassica campestris).
It is genetically modified (GMO) to improve oil quality and increase plant tolerance to herbicides Its
seeds are used to produce edible oil, suitable for consumption by humans and livestock.

Advantages and disadvantages of transgenic plants:

Advantanges: Gentically Modified plants have been useful in many ways such as:
 Genetic modification has: made crops more tolerant to abiotic stresses (cold, drought, salt, heat),
 Reduced reliance on chemical pesticides (pest-resistant crops).
 Helped to reducepost harvest losses.
 Increased efficiency of mineral usage by plants (this prevents early exhaustion of fertility of soil).
 Enhanced nutritional value of food, e.g., vitamin 'A' enriched rice.

Disadvantages: Genetically modified plants could pose biological and ecological risks like:
 Production of toxic or allergic metabolites.
 unexpected new susceptibilities to pathogens.
 Transmission of the new traits to related sexually compatible weed species.
 The ecosystems may be disturbed by dispersal, persistence or altered reaction to parasites, symbionts
or competitors.

Contacts: Suman Debnath, Ph-7687931008. E-Mail: [email protected], Class: XII (NEET) Page 2
BIO-SCOPE SUMAN DEBNATH (M.Sc in ZOOLOGY), B.Ed (Life Sciences)

Transgenic Animals
 Animals that have had their DNA manipulated to possess and express an extra (foreign) gene are known
as transgenic animals.
 In animals, transfection specifies the introduction of a DNA segment, either naked or integrated into a
vector, into an animal cell. The same phenomenon is known as transformation in all other organisms.
 Transgenic animals can be specifically designed to allow the study of how genes are regulated, and
how they affect the normal functions of the body and its development, e.g., study of complex factors
involved in growth such as insulin-like growth factor.
 Many transgenic animals are designed to increase our understanding of how genes contribute to the
development of disease.
 Transgenic animals that produce useful biological products can be created by the introduction of the
portion of DNA (or genes) which codes for a particular product such as human protein (a-1-antitrypsin)
used to treat emphysema. Similar attempts are being made for treatment of phenylketonuria (PKU)
and cystic fibrosis.

Transgenic cow: In 1997, the first transgenic cow, Rosie produced human protein-enriched milk (2.4
grams per litre). The milk contained the human alpha-lactalbumin and was nutritionally a more balanced
product for human babies than natural cow milk.

Transgenic mice are being developed for use in testing the safety of vaccine before they are used on
humans. Transgenic animals are made that carry genes which make them more sensitive to toxic substances
than non-transgenic animals. Toxicity testing in such animals will allow us to obtain results in less time.

Transgenic Fish: Gene transfers have been successful in various fish, such as common carp, rainbow trout,
Atlantic salmon, catfish, goldfish, zebra-fish etc. Genetically modified salmon was the first transgenic
animal obtained for food production.

Transgenic Rabbits are quite promising for gene farming or molecular farming, which aims at the
production of recoverable quantities of pharmaceutically or biologically important proteins encoded by the
transgenes.

Transgenic Goats are being evaluated as bioreactors. Some human genes have been introduced in goats and
their expression achieved in mammary tissues.

Transgenic sheep have been produced to achieve better growth and meat production. For example, human
genes for blood clotting factor IX and for α1antitrypsin have been transferred in sheep and expressed in
mammary tissue.

Biotechnological applications in medicine

The biotechnological processes have made great impact in the area of healthcare by mass production of safe
and more effective therapeutic drugs. This is known as medical biotechnology or red biotechnology.
Genetically engineered Insulin:
 Insulin for curing diabetes used to be extracted from pancreas of
slaughtered pigs and cattle.
 Insulin is made up of 51 amino acids arranged in two
polypeptide chains, (A chain and B chain).
 A having 21 amino acids and B with 30 amino acids that are
linked together by disulphide bridges.
 In mammals, including humans, insulin is synthesised as a pro-
hormone which contains an extra stretch called the C peptide.
 This C peptide is not present in the mature insulin and is
removed during maturation into insulin.

Contacts: Suman Debnath, Ph-7687931008. E-Mail: [email protected], Class: XII (NEET) Page 3
BIO-SCOPE SUMAN DEBNATH (M.Sc in ZOOLOGY), B.Ed (Life Sciences)

The main challenge for production of insulin using rDNA techniques was getting insulin assembled into a
mature form.1983, Eli Lilly an American company prepared two DNA sequences corresponding to A and B,
chains of human insulin and introduced them in plasmids of E. coli to produce insulin chains. Chains A and
B were produced separately, extracted and combined by creating disulfide bonds to form human insulin.

Gene therapy
It is a collection of methods that allows genes to be inserted into a person's cells and tissues to treat a
disease. Correction of a genetic defect involves delivery of a normal gene into the individual or embryo to
take over the function of and compensate for the non-functional gene.
 The first clinical gene therapy was given in 1990 to a 4-year old girl with adenosine deaminase
(ADA) deficiency.
 This enzyme is very important for the immune system to function.
 SCID is caused due to defect in the gene for the enzyme adenosine deaminase.
 In some children ADA deficiency can be cured by bone marrow trartsplantation. However, in others
it can be treated by enzyme replacement therapy in which functional ADA is given to the patient by
infiection. But in both approaches the patients are not completely cured.
 However, if the isolated gene from bone marrow cells producing ADA is introduced into cells at
early embryonic stages, it can be a permanent cure.

 Recombinant Vaccine: In case of many pathogens the gene encoding a critical antigen has been isolated
and expressed in bacteria or animals, and the recombinant protein so produced is used as a vaccine such
vaccines are called recombinant vaccines. A constituent polypeptide or a small part of a polypeptide
may be used as a vaccine such vaccines are known as subunit vaccines.

 Monoclonal antibody: It is a specific antibody produced from a normally short lived antigen-activated
B-cell that has been immortalized by hybridizing it with a myeloma cell by hybrid cell culture technique
known as hybridoma technology.

Molecular diagnosis
For effective treatment of a disease, early diagnosis and understanding its pathophysiology is very
important. Recombinant DNA technology, Polymerase Chain Reaction (PCR) and Enzyme Linked Immuno-
sorbent Assay (ELISA) are some of the techniques that serve the purpose of early diagnosis.

 Diagnosis by PCR technique: very low concentration of a bacteria or virus (at a time when the
symptoms of the disease are not yet visible) can be detected by amplification of their nucleic acid by
PCR. It is useful for detecting AIDS, Cancer and many other genetical disorders.
 DNA Probes: A single stranded DNA or RNA, tagged with a radioactive molecule (probe) is allowed to
hybridise to its complementary DNA in a clone of cells followed by detection using autoradiography. The
clone having the mutated gene will hence not appear on the photographic film, because the probe will not
have complementarity with the mutated gene.
 ELISA: (Enzyme Linked Immuno-sorbent Assay): ELISA is based on the principle of antigen-antibody
interaction. Infection by pathogen can be detected by the presence of antigens (proteins, glycoproteins,
etc.) or by detecting the antibodies synthesised against the pathogen.

Contacts: Suman Debnath, Ph-7687931008. E-Mail: [email protected], Class: XII (NEET) Page 4
BIO-SCOPE SUMAN DEBNATH (M.Sc in ZOOLOGY), B.Ed (Life Sciences)

Biosafety and Ethical Issues

It is mandatory to evaluate the morality of all human activities that might help or harm living organisms.

 GEAC: The Indian Government has setup an organization such as Genetic Engineering Approval
Committee (GEAC) which will make decision regarding the validity of GM research and the safety of
GM organisms for public services.

Bioethics includes rules of conduct that may be used to regulate our activities in relation to the biological
world. The main bioethical concerns pertaining to biotechnology are briefly mentioned as follows:

 Introduction of a transgene from one species into another species violates the 'integrity of species'.
 Biotechnology may pose unforeseen risks to the environment, including risk to biodiversity.
 Transfer of human genes into animals (and vice-versa) dilutes the concept of 'humanness'.
 When animals are used for production of pharmaceutical proteins, they are virtually reduced to the
status of a 'factory'.
 Use of animals in biotechnology causes great suffering to them.
 It is disrespectful to living beings, and only exploits them for the benefit of human beings.
 Scientists cannot rule out the possibility of other biological damage.
 It can accidentally create new infectious agents

The modification/usage of living organisms for public services (a food and medicine sources) has created
problems with patents and problems of biopiracy arised.

 Biopatent is the patent granted by certain companies for the products and technologies that make use of
the genetic materials, plants and other biological resources.
For example, an American company got patent rights on Basmati rice through the US Patent and
Trademark office. This allowed the company to sell a new variety of Basmati, in the US and abroad.

 Biopiracy refers to the use of the bioresources by multinational companies and other organizations
without proper authorisation from the countries and people concerned without compensatory payment.

Contacts: Suman Debnath, Ph-7687931008. E-Mail: [email protected], Class: XII (NEET) Page 5