Download Answer any EIGHT questions from Section A. Each question carries

APPLIED BIOTECHNOLOGY
Duration -3 Hrs
Max Marks: 70
Note:
1. Answer any EIGHT questions from Section A. Each question carries 5 marks.
2. Answer any THREE questions from Section B. Each question carries 10 marks.
Section A
Answer any EIGHT questions from Section A. Each question carries 5 marks.
1. Define biotechnology and mention about 3 ancient biotechnology processes
Ans. Biotechnology (sometimes shortened to "biotech") is a field of applied biology that involves the
use of living organisms and bioprocesses in engineering, technology, medicine and other fields requiring
bioproducts. Biotechnology also utilizes these products for manufacturing purpose. Modern use of similar
terms includes genetic engineering as well as cell and tissue culturetechnologies. The concept
encompasses a wide range of procedures (and history) for modifying living organisms according to
human purposes — going back to domestication of animals, cultivation of plants, and "improvements" to
these through breeding programs that employ artificial selection and hybridization. By comparison to
biotechnology, bioengineering is generally thought of as a related field with its emphasis more on higher
systems approaches (not necessarily altering or using biological materials directly) for interfacing with and
utilizing living things. TheUnited Nations Convention on Biological Diversity defines biotechnology as:[1]
"Any technological application that uses biological systems, living organisms, or derivatives thereof, to
make or modify products or processes for specific use."
In other terms: "Application of scientific and technical advances in life science to develop commercial
products" is biotechnology. Biotechnology draws on the pure biological sciences
(genetics, microbiology, animal cell culture, molecular biology, biochemistry,embryology, cell biology) and
in many instances is also dependent on knowledge and methods from outside the sphere of biology
(chemical engineering, bioprocess engineering, information technology, biorobotics). Conversely, modern
biological sciences (including even concepts such as molecular ecology) are intimately entwined and
dependent on the methods developed through biotechnology and what is commonly thought of as the life
sciences industry.
2. Write a brief note on CPCSEA guidelines
3. Write about equilibrium density-gradient centrifugation
Ans. centrifugation, also known as density gradient centrifugation or equilibrium sedimentation is a
technique used to separate molecules on the basis of buoyant density. (The word "isopycnic" means
"equal density".) Typically, a "self-generating" density gradient is established via equilibrium
sedimentation, and then analyte molecules concentrate as bands where the molecule density matches
that of the surrounding solution. To illustrate the process, consider the fractionation of nucleic acids such
as DNA. To begin the analysis, a mixture ofcaesium chloride and DNA is placed in a centrifuge for several
hours at high speed to generate a force of about 10^5 x g (earth's gravity). Caesium chloride is used
because at a concentration of 1.6 to 1.8 g/mL it is similar to the density of DNA. After some time a
gradient of the caesium ions is formed, caused by two opposing forces: diffusion and centrifugal force.
The sedimenting particles (caesium ions) will sediment away from the rotor, and become more
concentrated near the bottom of the tube. The diffusive force arises due to the concentration gradient of
solvated caesium chloride and is always directed towards the center of the rotor. The balance between
these two forces generates a stable density gradient in the caesium chloride solution, which is more
dense near the bottom of the tube, and less dense near the top.
The DNA molecules will then be separated based on the relative proportions of AT
(adenine and thymine base pairs) to GC (guanine andcytosine base pairs). An AT base pair has a lower
molecular weight than a GC base pair and therefore, for two DNA molecules of equal length, the one with
the greater proportion of AT base pairs will have a lower density, all other factors being equal. Different
types of nucleic acids will also be separated into bands, e.g. RNA is denser
than supercoiled plasmid DNA, which is denser than linear chromosomal DNA.
4. How do you increase the monoclonal antibody titre in a culture
5. Distinguish between mutualism and Parasitism?
Ans. Mutualism is the way two organisms of different species biologically interact in a relationship in
which each individual derives a fitness benefit (i.e., increased or improved reproductive output). Similar
interactions within a species are known as co-operation. Mutualism can be contrasted withinterspecific
competition, in which each species experiences reduced fitness, and exploitation, orparasitism, in which
one species benefits at the expense of the other. Mutualism is a type ofsymbiosis. Symbiosis is a broad
category, defined to include relationships that are mutualistic,parasitic, or commensal. Mutualism is only
one type.
A well known example of mutualism is the relationship between ungulates (such as cows)
andbacteria within their intestines. The ungulates benefit from the cellulase produced by the bacteria,
which facilitates digestion; the bacteria benefit from having a stable supply of nutrients in
the hostenvironment. Parasitism is a type of non mutual relationship between organisms of
different species where one organism, the parasite, benefits at the expense of the other, the host.
Traditionally parasitereferred to organisms with lifestages that needed more than one host (e.g. Taenia
solium). These are now called macroparasites (typically protozoa and helminths). The word parasite now
also refers to microparasites, which are typically smaller, such as viruses and bacteria, and can be directly
transmitted between hosts of the same species [1]. Examples of parasites include the plants mistletoe and
cuscuta, and organisms such as leeches.
Unlike predators, parasites are generally much smaller than their host; both are special cases
ofconsumer-resource interactions.[2] Parasites show a high degree of specialization, and reproduceat a
faster rate than their hosts. Classic examples of parasitism include interactions betweenvertebrate hosts
and diverse animals such as tapeworms, flukes, the Plasmodium species, andfleas.
Parasitism is differentiated from the parasitoid relationship, though not sharply, by the fact that parasitoids
generally kill or sterilise their hosts. Parasitoidism occurs in much the same variety of organisms that
parasitism does.
The harm and benefit in parasitic interactions concern the biological fitness of the organisms involved.
Parasites reduce host fitness in many ways, ranging from general or specialized pathology, such
as parasitic castration and impairment of secondary sex characteristics, to the modification of host
behaviour. Parasites increase their fitness by exploiting hosts for resources necessary for the parasite's
survival, e.g. food, water, heat, habitat, and genetic dispersion.
6. What is bioremediation?
Ans. Bioremediation is the use of microorganism metabolism to remove pollutants. Technologies can be
generally classified as in situ or ex situ.In situ bioremediation involves treating the contaminated material
at the site, while ex situ involves the removal of the contaminated material to be treated elsewhere. Some
examples of bioremediation technologies
are phytoremediation, bioventing, bioleaching, landfarming, bioreactor,composting, bioaugmentation, rhizo
filtration, and biostimulation.
Bioremediation can occur on its own (natural attenuation or intrinsic bioremediation) or can be spurred on
via the addition of fertilizers to increase the bioavailability within the medium (biostimulation). Recent
advancements have also proven successful via the addition of matched microbe strains to the medium to
enhance the resident microbe population's ability to break down contaminants. Microorganisms used to
perform the function of bioremediation are known as bioremediators.[1]
Not all contaminants, however, are easily treated by bioremediation using microorganisms. For
example, heavy metals such as cadmium andlead are not readily absorbed or captured by
microorganisms. The assimilation of metals such as mercury into the food chain may worsen
matters. Phytoremediation is useful in these circumstances because natural plants or transgenic
plants are able to bioaccumulate these toxins in their above-ground parts, which are then harvested for
removal.[2] The heavy metals in the harvested biomass may be further concentrated by incineration or
even recycled for industrial use.
The elimination of a wide range of pollutants and wastes from the environment requires increasing our
understanding of the relative importance of different pathways and regulatory networks to carbon flux in
particular environments and for particular compounds, and they will certainly accelerate the development
of bioremediation technologies and biotransformation processes.
7. How are PCBs biodegraded?
Ans. A printed circuit board, or PCB, is used to mechanically support and electrically connectelectronic
components using conductive pathways, tracks or signal traces etched fromcopper sheets laminated onto
a non-conductive substrate. It is also referred to as printed wiring board (PWB) or etched wiring board.
A PCB populated with electronic components is a printed circuit assembly (PCA), also known as
a printed circuit board assembly or PCB Assembly (PCBA). Printed circuit boards are used in virtually
all but the simplest commercially produced electronic devices.
Alternatives to PCBs include wire wrap and point-to-point construction. PCBs are often less expensive
and more reliable than these alternatives, though they require more layout effort and higher initial cost.
PCBs are much cheaper and faster for high-volume production since production and soldering of PCBs
can be done by automated equipment. Much of the electronics industry's PCB design, assembly, and
quality control needs are set by standards that are published by the IPC organization.
Hanson, described flat foil conductors laminated to an insulating board, in multiple layers. Thomas
Edison experimented with chemical methods of plating conductors onto linen paper in 1904. Arthur Berry
in 1913 patented a print-and-etch method in Britain, and in the United States Max Schoop obtained a
patent[1] to flame-spray metal onto a board through a patterned mask. Charles Durcase in 1927 patented
a method of electroplating circuit patterns. [2]
The Austrian Jewish engineer Paul Eisler invented the printed circuit while working in England around
1936 as part of a radio set. Around 1943 the USA began to use the technology on a large scale to
make proximity fuses for use in World War II [2]. After the war, in 1948, the USA released the invention for
commercial use. Printed circuits did not become commonplace in consumer electronics until the mid1950s, after the Auto-Sembly process was developed by the United States Army.
Before printed circuits (and for a while after their invention), point-to-point construction was used. For
prototypes, or small production runs,wire wrap or turret board can be more efficient. Predating the printed
circuit invention, and similar in spirit, was John Sargrove's 1936–1947 Electronic Circuit Making
Equipment (ECME) which sprayed metal onto a Bakelite plastic board. The ECME could produce 3 radios
per minute.
8. What are the characteristics of wastewater?
Ans. Wastewater is any water that has been adversely affected in quality by anthropogenic influence. It
comprises liquid waste discharged by domestic residences, commercial properties, industry, and/or
agriculture and can encompass a wide range of potential contaminants and concentrations. In the most
common usage, it refers to the municipal wastewater that contains a broad spectrum of contaminants
resulting from the mixing of wastewaters from different sources.
Sewage is correctly the subset of wastewater that is contaminated with feces or urine, but is often used to
mean any waste water. "Sewage" includes domestic, municipal, or industrial liquid waste
products disposed of, usually via a pipe or sewer or similar structure, sometimes in acesspool emptier.
The physical infrastructure, including pipes, pumps, screens, channels etc. used to convey sewage from
its origin to the point of eventual treatment or disposal is termed sewerage.
Wastewater or sewage can come from (text in brackets indicates likely inclusions or contaminants):

Human waste (fæces, used toilet paper or wipes, urine, or other bodily fluids), also known
asblackwater, usually from lavatories;

Cesspit leakage;

Septic tank discharge;

Sewage treatment plant discharge;

Washing water (personal, clothes, floors, dishes, etc.), also known as greywater or sullage;

Rainfall collected on roofs, yards, hard-standings, etc. (generally clean with traces of oils andfuel);

Groundwater infiltrated into sewage;

Surplus manufactured liquids from domestic sources (drinks, cooking oil, pesticides,lubricating
oil, paint, cleaning liquids, etc.);

Urban rainfall runoff from roads, carparks, roofs, sidewalks, or pavements (contains oils, animal
fæces, litter, fuel or rubber residues, metals from vehicle exhausts, etc.);
9. Write about the physical properties and chemical composition of culture media
10. Discuss polymerase chain reaction?
Ans. The polymerase chain reaction (PCR) is a scientific technique in molecular biology to amplify a
single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to
millions of copies of a particular DNA sequence.
Developed in 1983 by Kary Mullis,[1] PCR is now a common and often indispensable technique used in
medical and biological research labs for a variety of applications.[2][3] These include DNA
cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; the diagnosis of hereditary
diseases; the identification of genetic fingerprints (used in forensic sciences and paternity testing); and the
detection and diagnosis of infectious diseases. In 1993, Mullis was awarded the Nobel Prize in
Chemistry along with Michael Smith for his work on PCR.[4]
The method relies on thermal cycling, consisting of cycles of repeated heating and cooling of the reaction
for DNA melting and enzymatic replication of the DNA.Primers (short DNA fragments) containing
sequences complementary to the target region along with a DNA polymerase (after which the method is
named) are key components to enable selective and repeated amplification. As PCR progresses, the
DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the
DNA template is exponentially amplified. PCR can be extensively modified to perform a wide array
of genetic manipulations.
Almost all PCR applications employ a heat-stable DNA polymerase, such as Taq polymerase, an enzyme
originally isolated from the bacterium Thermus aquaticus. This DNA polymerase enzymatically assembles
a new DNA strand from DNA building-blocks, thenucleotides, by using single-stranded DNA as a template
and DNA oligonucleotides (also called DNA primers), which are required for initiation of DNA synthesis.
The vast majority of PCR methods use thermal cycling, i.e., alternately heating and cooling the PCR
sample to a defined series of temperature steps. These thermal cycling steps are necessary first to
physically separate the two strands in a DNA double helix at a high temperature in a process called DNA
melting. At a lower temperature, each strand is then used as the template in DNA synthesis by the DNA
polymerase to selectively amplify the target DNA. The selectivity of PCR results from the use
of primers that arecomplementary to the DNA region targeted for amplification under specific thermal
cycling conditions.
Answer any THREE questions from Section B. Each question carries 10 marks.
Section B
11. Debate reproductive cloning in animals versus humans
Ans. Human–animal communication is easily observed in everyday life. The interactions between pets
and their owners, for example, reflect a form of spoken, while not necessarily verbal dialogue. A dog
being scolded does not need to understand every word of its admonishment, but is able to grasp the
message by interpreting cues such as the owner's stance, tone of voice, and body language. This
communication is two-way, as owners can learn to discern the subtle differences between barks and
meows … one hardly has to be a professional animal trainer to tell the difference between the bark of an
angry dog defending its home and the happy bark of the same animal while playing. Communication
(often nonverbal) is also significant in equestrian activities such as dressage.
[edit]Word
repetition in birds
Although the word repetition skills observed in some birds (most famously parrots) should not be mistaken
for lingual communication, this tendency has nonetheless influenced fictional portrayals of animal
communication, as sentient talking parrots and similar birds are common in children's fiction, such as the
talking, loud-mouth parrot Iago of Disney's Aladdin. Bruce Thomas Boehner's book Parrot Culture: Our
2,500-Year-Long Fascination with the World's Most Talkative Bird explores this issue thoroughly.
[edit]The
next level: language
Achieving a deeper level of communication between animals and humans has long been a goal of
science. Perhaps the most famous example of recent decades has been Koko, a gorilla who is
supposedly able to communicate with humans using a system based onAmerican Sign Language with a
"vocabulary" of over 1000 words.
[edit]John
Lilly and cetacean communication
In the 1960s, John Lilly, M.D., prolific writer and explorer of consciousness via the isolation tank (his
invention), and contemporary and associate of Timothy Leary, began experiments in the Virgin
Islands aiming to establish meaningful communication between humans and thebottlenose
dolphin (Tursiops truncatus). Lilly financed, mostly personally, a human-dolphin cohabitat, a house on the
ocean's shore that contained an area that was partially flooded and allowed a human and dolphin to live
together in the same space, sharing meals, play, language lessons, and even sleep.
Two experiments of this sort are explained in detail in Lilly's popular books (see John Lilly for
bibliography). The first experiment was more of a test run to check psychological and other strains on the
human and cetacean participants, determining the extent of the need for other human contact, dry
clothing, time alone, and so on. Despite tensions after several weeks, the experimenter, Margaret C.
Howe, agreed to a two-and-a-half month experiment, living isolated with 'Peter' dolphin.
12. List the gene delivery methods with special mention to microinjection
Ans. Microinjection refers to the process of using a glass micropipette to insert substances at
a microscopic or borderline macroscopic level into a single living cell. It is a simple mechanical process in
which a needle roughly 0.5 to 5 micrometers in diameter penetrates the cell membrane and/or the nuclear
envelope. The desired contents are then injected into the desired sub-cellular compartment and the
needle is removed. Microinjection is normally performed under a specialized optical microscope setup
called a micromanipulator. The process is frequently used as a vector in genetic
engineering and transgenics to insert genetic material into a single cell. Microinjection can also be used in
the cloning of organisms, and in the study of cell biology and viruses. Microcapillary and microscopic
devices are used to deliver DNA into a protoplast.
[edit]Examples

Microinjection is used as a vector in transgenic plant production.

Microinjection of genes into fertilized eggs is a common vector used in the production of higher forms
of transgenic animals.

Microinjection of a gene knockdown reagent such as a morpholino oligo into eggs or early zygotes is
commonly used to probe the function of a gene during development of embryos.
13. Define the usage of progesterone, prostoglandin and PMSG
Mention the importance of marker genes in DNA transfection
Ans. Transfection is the process of deliberately introducing nucleic acids into cells. The term is used
notably for non-viral methods in eukaryotic cells[1]. It may also refer to other methods and cell types,
although other terms are preferred: "transformation" is more often used to describe non-viral DNA transfer
in bacteria, non-animal eukaryotic cells and plant cells - a distinctive sense of transformation refers to
spontaneous genetic modifications (mutations to cancerous cells (carcinogenesis), or under stress (UV
irradiation)). Transduction is often used to describe virus-mediated DNA transfer. The word transfection is
a blend of trans- and infection.
Genetic material (such as supercoiled plasmid DNA or siRNA constructs), or even proteins such
as antibodies, may be transfected.
Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane, to
allow the uptake of material. Transfection can be carried out using calcium phosphate, by electroporation,
or by mixing a cationic lipid with the material to produceliposomes, which fuse with the cell membrane and
deposit their cargo inside.
Transfection can result in unexpected morphologies and abnormalities in target cells.
The meaning of the term has evolved.[2] The original meaning of transfection was "infection
by transformation," i.e., introduction of DNA (or RNA) from a prokaryote-infecting virus
or bacteriophage into cells, resulting in an infection. Because the term transformation had
another sense in animal cell biology (a genetic change allowing long-term propagation in
culture, or acquisition of properties typical of cancer cells), the term transfection acquired, for
animal cells, its present meaning of a change in cell properties caused by introduction of
DNA.
14. Why cell Ca2+ is an inexpensive but cAMP is an expensive second messenger?