This is one of the major chapters for the examination NEET. A few questions are expected from this chapter and carry more weight as per the NEET syllabus.

1. NEET/NCERT/NTA
BIOLOGY-NOTE
BIOTECHNOLOGY
PROCESS AND APPLICATION
DR.ASHOKAN KANNARATH

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CHAPTER -3
BIOTECHNOLOGY: PROCESS AND APPLICATION
Biotechnology is a branch of biology which deals with techniques of using live organisms or
enzymes from organisms to produce products and processes useful to humans.
Oldest form biotechnology
Making curd, bread or wine, which is all microbe-mediated processes, could also be thought as a
form of biotechnology. The process of fermentation also we use in the production of wine,
alcoholic beverages.
Modern form of biotechnology
Today the term biotechnology is used in a restricted sense to refer such a processes which use
genetically modified organisms to achieve the same products, as in older biotechnology, on a larger
scale. Further, many other processes/techniques are also included under biotechnology. For
example, in vitro fertilization leading to a test tube baby, synthesizing a gene and suing it,
developing a DNA vaccine or correcting a defective gene, are all part of biotechnology. The
European Federation of Biotechnology (EFB) has given a definition of biotechnology that
encompasses both traditional view and modern molecular biotechnology. The definition given by
EFB is as follows: ―The integration of natural science and organisms, cells, parts thereof, and
molecular analogues for products services‖
Recombinant DNA technology or Genetic engineering
Variation is the raw material of evolution. Variation is not produced in asexual reproduction, but it
is possible in sexual reproduction. The variation produced in the sexual reproduction may be
desirable or undesirable and hence it is not predetermined. To produce a predetermined desirable
change in the genome is genetic engineering or recombinant DNA technology. Genetic engineering
or rDNA technology involves a manipulation of the genetic material towards a desired end and in
predetermined way.
Significance: It by pass the restriction in the gene transfer mechanism between unrelated
organisms.
Difference between genetic recombination and genetic engineering is that recombination
occurs by natural process while genetic engineering is the purposeful manipulation or engineering
of genes and to produce desirable change in the gene.
The recombinant DNA technology was first proposed by Peter Lobban. The present day
rDNA technology was flourished after Cohen and Herbert Boyer in 1972.He successfully link a
gene coding for antibiotic resistance with a native plasmid of Salmonella typhimurium with the
vector plasmid and then cloning it in E.coli. Biotechnology can be defined as ―the addition,
removal, replacement or repair of a part of genetic material resulting in to the change of phenotype
of an organism is called genetic engineering‖

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rDNA technology: Recombinant DNA ethnology is the technique of manipulating the genome of a
cell or organism so as to change the phenotype desirably.
Basic steps involved in rDNA technology
1. Isolating genomic DNA of a ―donor‖. Donor is the cell or organism from which the required gene
is taken.
2. Fragmenting this DNA using ―molecular scissor‖, Restriction endonuclease.
3. Screening the fragments for desired gene.
4. Inserting the fragments with desired gene into a ―cloning vector‖(a plasmid, cosmid or a phage
DNA).
5. Introducing the recombinant vector into a competent host cell.
6. Culturing these cells to obtain multiple copies or clones of desired fragments of DNA.
7. Using these copies to ‗Transform‘ suitable host cells so as to express the desired gene.

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Tools used in genetic engineering
The main tools used in genetic engineering are as follows:
1. Enzymes: The different enzymes used in genetic engineering are the following:
A. Cleaving enzyme: These enzymes are used to break DNA molecules. They are of three
types:
1.Exonuclease: They remove nucleotides from the terminal ends, either 5` or 3` of DNA in
one strand of double stranded DNA
2.Endonuclease: They make cuts at specific position within the DNA. This enzymes cut
only in one strand of the double stranded DNA.
3.Restriction endonuclease: Restriction endonucleases are called ―molecular scissors‖ or
―biological scissors‖. They recognize and cut double stranded DNA at specific points called
palindromic sequences. Palindromic sequences are the one which read same on both the strand in
5`→3` direction. Same as true about 3`→5` direction.
↓
5`−G AATTC−3`
3`−CTTAA G−5`
↑
Restriction endonuclease was discovered by Arber in 1963 in bacteria. The firs restriction
endonuclease discovered is EcoR1 found in colon bacterium Escherichia coli.
Types of REN
Three main types of REN are present-Type-I, Type-II and Type-III. Type-II REN is commonly
used in rDNA technology because it can be used in vitro to identify and cleave within specific DNA
sequence usually having 4-8 nucleotides. More than 350 different type-II endonuclease with 100
different recognition sequences are known.
Nomenclature of Restriction endonuclease
REN are named by certain standard procedure, the first letter of the enzymes indicates the genus
name followed by the first two letters of the species name, and then comes the strain and finally
Roman numerals indicating the order of discovery.
E.g. EcoRI- E -for Echeratia
co- for coli
R-for Ry13 strain
I-For First endonuclase discovered.
Hind III- H- for haemophilus
In-influenza d- for
Rd strain
III-third endonuclease discovered.
Sticky ends
Sticky ends are overhang/stretch of single stranded portion generated as a result of cut by restriction
endonuclease into two fragments. They are named so because they form hydrogen bonds with their

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complementary counter parts. The stickiness of the end facilitates the action of enzyme DNA ligase.
All the fragments cut by a restriction endonuclease will have the same sticky ends. Sticky ends have
complementary base. So the y could join.
Some of the restriction endonuclease and their sources are the following
Cutting of DNA by restriction endonuclease result in mixture of DNA strands of varied length. This
is called restriction digestion. DNA of various lengths is isolated on a gel plate under electric field
by technique called gel electrophoresis. The main dye in agarose gel electrophoresis is ethidium
bromide. Restriction digest is subjected to electrophoresis to separate DNA on the basis of length.
DNA are (-) vely charged and move towards (+) vely charged anode in a gel with definite pore size.
Smaller DNA moves faster and farthest.
Elusion: It is the extraction of DNA bands out of agarose gel and visualizing it by staining with
ethidium bromide compound followed by exposure to UV radiation. They are visible as arrange of
colour bands.
DNA ligase
In 1969 Har Govind Khorana discovered DNA ligase in T-4 bacteriophage. They join two
individual fragments of double stranded DNA by forming phosphodiester bonds between them.
Thus they help in sealing gaps in DNA fragments. Therefore they acts as a molecular glue. The
enzyme used most commonly in rDNA technology is T4DNA ligase.

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Reverse transcriptase
Enzyme necessary for the conversion of RNA genetic material into DNA.
Alkaline Phosphotase
These enzymes release phosphate group from the 5 end of linearised circular DNA to check its
recircularization.
DNA polymerase
A DNA polymerase is a cellular or viral polymerase enzyme that synthesizes DNA molecules from
their nucleotide building blocks.
Vectors
Vectors are carrier of DNA molecule to which the fragments of desired DNA are attached and
carried to required site. The vector may be plasmid, transposons, and bacteriophage, DNA from
plants and animals or artificial DNA, lambda phages, baculovirus is useful in insects, Ti plasmid is
for plants and YCA (Yeats artificial chromosomes) is for yeast cells.
Characters of vector
1. It must have low molecular weight
2. It must be able to replicate inside the host.
3. It must have a replication origin. Replication origin is a sequence element which is
recognized by the host cell‘s replication machinery.
4. Thus cloning vector can be amplified in the host cell, and DNA segment inserted into the
vector amplified along with vector.
5. Vector also must have restriction endonuclease recognition sites and some marker gene
which will provide resistance to antibiotics and express in the host cell.
6. Marker in the vector helps to identify transformed bacterial colony.
Process of rDNA technology
DNA taken from both the sources are fragmented by restriction endonuclease.
The restriction endonuclease cuts both molecules at the specific site.
The ends of the cut have an overhanging piece of single-stranded DNA called ―Sticky ends‖. These
sticky ends are able to base pair with any DNA fragment that contains the complementary sticky
end. Enzyme DNA ligase is used to covalently link the two strands into a molecule of rDNA. This
rDNA needs to be replicated many times. Cloning can be done in vitro by PCR or in vivo by using
E.coli or yeast or mammalian tissue culture cell.
Products made by rDNA technology
1. Blood protein: Erythropoietin, Factor VII, VIII, IX, tissue plasminogen activator (TPA),
urokinase.
2. Human hormone, epidermal growth factor, FSH, insulin, nerve growth factor, relaxin,
somatotropin etc.
3. Immune modulators, ά-interferon, β-interferon, colony stimulating factor, lysozyme, tumor
necrosis factor.
4. Vaccines against cytomegaly virus, hepatitis –B virus, measles virus and rabies virus.

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Transposes
Transposes are segments of DNA that can move around to different positions in the genome of a
single cell. In the process, they may
1. Cause mutations
2. Increase (or decrease) the amount of DNA in the genome of the cell, and if the cell is the
precursor of a gamete, in the genomes of any descendants. These mobile segments of DNA are
sometimes called "jumping genes".
There are two distinct types:
1. Class I transposons. These are retrotransposons that
1. first transcribe the DNA into RNA and then
2. Use reverse transcriptase to make a DNA copy of the RNA to insert in a new
location.
2. Class II transposons. These consist of DNA that moves directly from place to place.
Class II Transposons
Class II transposons move by a "cut and paste" process: the transposon is cut out of its location (like
command/control-X on your computer) and inserted into a new location (command/control-V).This
process requires an enzyme — a transposase — that is encoded within some of these transposons.
Transposase binds to:
1. Both the ends of the transposon, which consist of inverted repeats; that is, identical
sequences reading in opposite directions.
2. A sequence of DNA that makes up the target site. Some transposases require a specific
sequence as their target site; others can insert the transposon anywhere in the genome.
The DNA at the target site is cut in an offset manner (like the "sticky ends" produced by some
restriction enzymes [Examples]). After the transposon is ligated to the host DNA, the gaps are
filled in by Watson- Crick base pairing. This creates identical direct repeats at each end of the
transposon. Often transposons lose their gene for transposase. But as long as somewhere in the cell
there is a transposon that can synthesize the enzyme, their inverted repeats are recognized and they,
too, can be moved to a new location.
Plasmids

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Plasmids are small, circular DNA molecules present in bacterial cells in addition to the
chromosomal DNA and that replicates autonomously. The term plasmid was first introduced by
Joshua Lederberg in 1952 (American Molecular biologist). The size of plasmid varies from 1 -1,000
kilo base pairs (kbp). Genes from one organism may be added to the plasmids of a bacterium and
transfer them to another. They retain their characters after combining with the DNA of other
organisms. Not being a part of the main genome, they can be easily isolated and transferred. These
features make the plasmids most suitable for use as a vehicle DNA. A plasmid having DNA of
another organism (lambda phage) integrated with it is known as cosmid, also recombinant plasmid
or hybrid plasmid. Cosmids are picked up by or can be introduced into plasmid free bacteria.
The plasmids are designated by a lower case P (p) followed by the first letter/s of
researcher/s names and the numerical number given by the workers. E.g. pBR322 is as plasmid
discovered by Bolivar and Rodriguez, who designated it as 322. Some plasmids are given name of
the place where they are discovered e.g. pUC is plasmid from University of California.
Bacteriophage: Bacteriophage is a virus that infects bacteria.
Structure
Bacteriophage consists of an outer protein capsid enclosing the genetic material. The genetic
material can be ssRNA, dsRNA, ssDNA or dsDNA. It may be either circular or linear.
Use of bacteriophage in rDNA
The cloning of single gene is carried out by using plasmid. But cloning of larger pieces of DNA
plasmids is not suitable because larger inserts increase the size of the plasmid making the
transformation difficult. Commonly used bacteriophages as cloning vectors are M13 and lambda
phage which infect E.coli.
Phage Lambda (λ)a vector
DNA of phage lambda 48.5 kb in length. The ‗cose‘ cohesive allow DNA to be circularized in the
host cell. For cloning of large DNA fragments up to 20kbp DNA of phage is removed and replaced
by the DNA with desired gene. The recombinant DNA is then packaged within viral particles in
vitro and these are allowed to infect bacterial cells which have been cultured. Once inside the
bacterial cells, the recombinant viral DNA starts replicating.

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Replication of bacteriophage (Lytic cycle)
Inside the specific host/ bacterial cell the lytic cycle involves the following steps
1. Attachment: Bacteriophage attach to specific receptors on the surface of bacteria.
2. Penetration: After contact the tail fibers bring the base plate closer to the surface of the cell.
Then the tail contracts, injecting genetic material (DNA) through the bacterial membrane. The
capsid protein coat remain outside and is called ‗ghost‘.
3. Synthesis of proteins and nucleic acids: the phage DNA forced the bacterial metabolic
machinery to synthesis viral DNA and proteins.
4. Viron assembly: The base plates are assembled with tails first. The heads-capsid is constructed
separately and then is joined with the tails. The DNA is packed efficiently within the head. The
whole process complete within 15 minutes.
5. Release of virons: Phages are released via lysis of cells with the help of endolysin enzyme.
Released virons are capable of infecting new cells.
Bacteriophage –Lytic cycle
Passenger DNA
It is the DNA that is transferred from one organism into another by combining it with the vehicle
DNA. Three types of DNA are used as passengers: complimentary, synthetic and random
Complementary DNA (cDNA)
It is the DNA synthesized on RNA template with the help of the enzyme reverse transcriptase and

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necessary nucleotides. The DNA strand is isolated from the hybrid RNA-DNA complex by using
alkaline phosphatase enzyme. A complementary DNA strand is then synthesized on the isolated
single-stranded DNA template with the help of DNA polymerase. The cDNA double helix so
obtained can be joined with the vehicle DNA for introduction into a new host cell.
Synthetic DNA (sDNA)
It is the DNA synthesized with the help of DNA polymerases on the DNA template or from free
deoxybironucleotides without a template.
Production of restriction fragments
It was found that Hind II always cut DNA molecules at a particular point by recognizing a specific
sequence of six base pairs. This specific base sequence is known as the recognition sequence for
Hind II. Besides Hind II, today we know more than 900 restriction enzymes that have been
isolated from over 230 strains of bacteria each of which recognize different recognition sequences.
The convention for naming these enzymes is the first letter of the name comes from the genes and
the second two letters come from the species of the prokaryotic cell from which they were isolated,
e.g., EcoRI comes from Escherichia coli RY 13. nEoI h etr‗‗ sdrvdfo h aeo tan oa ubr following
the names indicate the order in which the enzymes were isolated from that strain of bacteria.
Restriction enzymes belong to a larger class of enzymes called nucleases. These are of two kinds;
exonucleases and endonucleases. Exonucleases remove nucleotides from the ends of the DNA
whereas; endonucleases make cuts at specific positions within the DNA. Each restriction
endonuclease functions y‗npcig‗telnt faDA sequence. Once it finds its specific recognition
sequence, it will bind to the DNA and cut each of the two strands of the double helix at specific
points in their sugar-phosphate backbones). Each restriction endonuclease recognizes a specific
palindromic nucleotide sequences in the DNA.
Palindromes are groups of letters that form the same words when read both forward and backward,
e.g., MLYLM. As against a word palindrome where the same word is read in both directions, the
palindrome in DNA is a sequence of base pairs that reads same on the two strands when orientation
of reading is kept the same. For example, the following sequences reads the same on the two
strands in 5`-3` direction. This is also true if read in the 3'-5' direction.
5' —— GAATTC —— 3'
3' —— CTTAAG —— 5'

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Restriction enzymes cut the strand of DNA a little away from the centre of the palindrome sites, but
between the same two bases on the opposite strands. This leaves single stranded portions at the
ends. There are overhanging stretches called sticky ends on each strand. These are named so
because they form hydrogen bonds with their complementary cut counterparts. This stickiness of
the ends facilitates the action of the enzyme DNA ligase. Restriction endonucleases are used in
genetic engineering to form recombinant molecules of DN. Which are composed of DNA from
different sources/ genomes? When cut by the same restriction enzyme, the resultant DNA fragments
have the same kind of ‗sticky ends‘, these can be joined together (end-to-end) using DNA ligases.
Preparing a DNA clone library
DNA clone library or Gene library is a collection of all cloned genes of an organism. It is of two
types
1. Genomic librray
2. cDNA librray
1. Genomic library
Genomic library is a collection of all clones that represent a complete set of genes (DNA
sequences) of an organism. It may be virus, bacteria, a plant or an animal.
Constructing genome library
1. Isolate entire genome.
2. Cut the genome into fragments of equal size by restriction endonuclease enzyme.
3. These fragments are then inserted into the cloning vectors.
4. The recombinant vectors are then transferred to suitable organism such as bacteria, yeast or
a virus.
5. These organisms are then cultured to produce their clones and stored.
6. Example: Genomic library of E.coli, the number of fragments of DNA required to construct
genomic library is 1,500, yeast-4,600, drosophila-48,000 and man-8,00,000.
2.cDNA library
cDNA is a copy of DNA which is produced by using m-RNA by the process of reverse
transcription. It is done with the help of the enzyme reverse transcriptase. This process is also
called as ‗teminism‘ since Temin and Baltimore discovered it. The library constructed by using
cDNA is called cDNA library.
In eukaryotes there are coding and non-coding sequences in the genes. The mRNA carying
only the trasncribed coding sequences in translated into protein. When mRNA of different cells,
tissues and organs at different time and phases in the life cycle of an organsism are isolated and
using revrse transcriptase enzyme converted into cDNA. A librray having cDNA for each type is
constructed by inserting into a suitable vector and then cloning it in a proper host like E.coli.
Polymer chain Reaction (PCR) or Gene amplification
The purpose of a PCR (Polymerase Chain Reaction) is to make a huge number of copies of a gene.
It was first developed in 1983 by Kary Mullis (Awarded Nobel prize for Chemistry in 1993). This
is necessary to have enough starting template for sequencing.
Basic requirement of PCR
1. A DNA segment (100-35,000 bp) to be amplified.

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2. Primer (forward and reverse) which are synthetic oligo-nucleotides of 17-30 nucleotides.
They are complementary to the sequence present on the desired DNA segments.
3. Four types of deoxyribonulceotides (dATP, dCTP, dGTP, dTTP). They are collectively
called dNTPs.
4. A thermo stable DNA polymerase that can withstand up to 94º C (Tag polymerase from
Thermus aquaticus)
5. An ideal PCR machine.
The cycling reactions:
There are three major steps in a PCR, which are repeated for 30 or 40 cycles. This is done on an
automated cycler, which can heat and cool the tubes with the reaction mixture in a very short time.
1. Denaturation at 91°C:
During the denaturation, the double strand melts open to single stranded DNA, all enzymatic
reactions stop (for example: the extension from a previous cycle).
2. Annealing at 55°C:
The primers are jiggling around, caused by the Brownian motion. Ionic bonds are constantly formed
and broken between the single stranded primer and the single stranded template. The more stable
bonds last a little bit longer (primers that fit exactly) and on that little piece of double stranded
DNA (template and primer), the polymerase can attach and starts copying the template. Once there
are a few bases built in, the ionic bond is so strong between the template and the primer, that it does
not break anymore.
3. Polymerization or Extension at 72°C:
This is the ideal working temperature for the polymerase. Tag polymerase adds dNTPs behind the
primer on the ssDNA.
Polymerase chain reaction

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These three steps constitute one cycle of the reaction. The process is carried out 28-30 cycles. Each
cycle of PCR takes about 3-5 minutes. As PCR progresses the DNA generated is itself used as a
template for replication, thus setting a chain reaction in which DNA template is exponentially
amplified.
Because both strands are copied during PCR, there is an exponential increase of the
number of copies of the gene. Suppose there is only one copy of the wanted gene before the cycling
starts, after one cycle, there will be 2 copies, after two cycles, there will be 4 copies, and three
cycles will result in 8 copies and so on.
Application of biotechnology in Agriculture-BT crops
Bacillus thuringiensis (or Bt) is a Gram-positive, soil-dwelling bacterium, commonly used as a
biological pesticide. Bt toxin proteins occur as inactive protoxins. When an insect ingests it, due to
the alkaline PH
of the gut, it gets converted into the active alkaline pH
of the gut; it gets converted
into the active form. The activated toxin cause swelling and paralyze the digestive system, and the
infected insect stops feeding within hours. Bt-affected insects generally die from starvation, which
can take several days. The gene for Bt toxin is called ‗cry‘ gene.bt toxin has been cloned and
introduced in many plants to provide resistance to insects without the need of insecticides. Bt cotton
is commercially available to control the disease of cotton ball. Other examples re Bt corn, rice,
tomato, potato and soybean.
Disadvantages
1. Bt is susceptible to degradation by sunlight. Most formulations persist on foliage less than a week
following application. Some of the newer strains developed for leaf beetle control become
ineffective in about 24 hours.
2. As strictly a stomach poison insecticide, Bt must be eaten to be effective
3. Its usefulness against pests that are susceptible to Bt but rarely have an opportunity to eat it in
field use, such as codling moth or corn earworm that tunnel into plants.
4. Since Bt does not kill rapidly, users may incorrectly assume that it is ineffective a day or two
after treatment.
5. Bt-based products tend to have a shorter shelf life than other insecticides.
Advantages
1. The specific activity of Bt generally is considered highly beneficial.
2. Unlike most insecticides, Bt insecticides do not have a broad spectrum of activity, so they do
not kill beneficial insects.
3. Perhaps the major advantage is that Bt is essentially nontoxic to people, pets and wildlife.
4. This high margin of safety recommends its use on food Crops or in other sensitive sites where
pesticide use can cause adverse effects.
Agrobacterium tumifaciens
It is a soil bacterium which causes crown gall tumor in dicotyledonous plants. A gall producing
gene (T DNA) occurs in a large plasmid called tumor inducing plasmid or Ti plasmid. The bacteria
insert Ti plasmids into nuclear genome of infected plants. This property of the bacterium utilized to

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transfer genes in higher plants through Ti-plasmids. Today cry gene from Bacillus thurengensis, Nif
genes (N2 fixing gene) from Rhizobium is cloned inside A .tumifaciens and then transferred into
other plants. Flavr savr tomato is developed by introducing an antisense gene that retard ripening.
It produces less cell wall degrading enzyme polygalactouranase. Hence the tomato has longer shelf
life. Golden rice is genetically engineered rice with greater pro-vitamin A content.
Bio-safety issues
Bio-safety is the prevention of large-scale loss of biological integrity, focusing both on ecology and
human health. Genetic modification of organisms can have unpredictable effects when such
organisms are introduced into ecosystem. For example, cross pollination between GM plants and
wild plants will lead to contamination of gene pools of wild varieties. Consumption of GM food
may develop allergies. GM microbes may escape from laboratories and will be hazardous.
Therefore manipulations of living organisms need regulations. Indian Government has set up
genetic engineering Approval committee (GEAC), which will take decisions regarding the validity
of GM research and safety of introducing GM product for public services.
Biopatent
A biopatent is a patent granted by the Government to the inventor for biological entities and for
products obtained from them.
1. The biopatens are awarded for genetically modified (GM) strains of microorganisms, plants,
animals, cell lines or DNA sequences or protein encoded by them or different industrial process
or various biotechnological procedures and products.
2. In the absence of patent the incentive for the research and development become reduced.
3. Patent may accelerate progress in biotechnology.
4. The profits are received due to legal protection of new products.
5. Patents are the most valuable type of intellectual property protection.
6. It gives the owner exclusive rights to market a product or invention and their by earn profits.

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7. They encourage technological innovations, investments and the development of the beneficial
products.
8. There are two different aspects of bio patents:
A. The biopatent should be granted carefully, fairly and impartially.
B. There are legal, ethical, economic and social implications of modern biotechnology which must
be thoroughly examined.
Biopiracy
The unlawful exploitation of the biopattented bioresources and also biopatening of bioresources of
other nation without proper permission of the concerned nation is considered as biopiracy.
The basic requirements for proper functioning of biopatents are the following:
1. The nation should be rich in biodiversity to provide ample bioresources.
2. The people living there should have proper traditional knowledge.
3. The nation should have sufficient financial resources for different experimentations.
It observed that the developed nation have very sound financial set up but poor bio-resources and
traditional knowledge, while the developing nations have good bio-resources and traditional
knowledge. This leads to biopiracy. Thus biopiracy is an opposite meaning of bioptent. A Texas
based company got patent right on Basmati rice through US patent and Trademark office. This
allowed the company to sell a new variety of basmati- Taximati. , in US and abroad. This new
variety is derived by crossing Indian basmati rice with semi dwarf variety and claimed as an
invention or a new variety. Thus it i s a case of Biopiracy. Patents on turmeric, margosa and many
other Indian medicinal plants are also attempted to Biopiracy.
QUESTIONS
1. What is biotechnology? Name a few fields in which biotechnology is applied traditionally.
2. Name a few fields in which biotechnology involves.
3. Name a few techniques in which biotechnology involves.
4. What is EFB?
5. What is biotechnology according to EFB?
6. What are the differences between sexual reproduction and asexual reproduction?
7. What is genetic engineering?
8. What is the basic aim of genetic engineering?
9. What is the difference between genetic engineering and genetic recombination?
10. Who proposed rDNA technology at first?
11. What is the contribution of Cohen and Boyer in the field of rDNA technology?
12. What is rDNA technology? Name any four scientist associated with it.
13. Explain basic steps involved in rDNA technology process.
14. What are the different tools involved in rDNA technology?
15. What is biological/chemical or molecular scissor or scalpel or knife?
16. What is vector in rDNA technology? Write briefly on it.
17. What are the criteria for vector in rDNA technology?

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18. What is marker in vector? Give its significance.
19. What is transformation?
20. What is Transfection?
21. What is transduction?
22. Name vector for
a) Bacteria
b) Insect
c) Plants
d) Yeast
23. Explain the process of rDNA technology
24. How can we do cloning in vivo and in vitro?
25. What is “sticky ends” in DNA?
26. Give a list of therapeutic products made by rDNA techniques.
27. What are transposons or jumping genes?
28. What is retro transposon? Explain
29. What is DNA transposon? Explain
30. What is plasmid? Write a note on plasmids.
31. What is Cosmid? Give its significance.
32. Write briefly on naming a plasmid.
33. What is Bacteriophage?
34. What is the genetic material in Bacteriophage?
35. What is the significance of Bacteriophage DNA as vector?
36. What is the genetic material of M13 and Lambda Bacteriophage?
37. Write a note on Phage lambda as vector.
38. What is Bacteriophage? Explain the mechanism of replication of Bacteriophage or lytic cycle.
39. What is ‘Ghost’ in Bacteriophage?
40. What is a restriction fragment? What is restriction endonuclease? Who discovered it?
41. Write a note on types of REN
42. Write briefly on nomenclature of REN
43. What is palindrome sequence? Give an example diagrammatically.
44. Write a note on cleavage pattern by REN
45. What is genomic library? Explain
46. What is cDNA library? Explain.
47. What is PCR? Give its significances.
48. What are the basic requirements for PCR techniques?
49. Name a thermo stable DNA polymerase and its source.
50. Explain steps involved in PCR with a suitable diagram.
51. What are transgenic plants? Give its advantages.
52. Write briefly on Bacillus thurengensis (Bt) as biopsied.
53. Write briefly on Agrobacterium tumefaciens. How it help in making genetically modified crops?

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54. Name any two GM plants and its benefits.
55. What is biosafety? Write a note on it
56. What is biopiracy? Write a note on it.
57. What is biopatent? Write briefly on it.

18. https://biol4u.com