Important Questions for Class 12

Biology
Chapter 11 - Biotechnology Principles and Processes

Very Short Answer Questions 1 Mark

1. A restriction enzyme digests DNA into fragments. Name the technique

used to check the progression of this enzyme and separate DNA fragments.
Ans: Gel electrophoresis is the technique in which DNA fragments are separated
after the digestion of DNA by a restriction enzyme that results in the formation
of fragments.

2. Name two commonly used vectors in genetic engineering.

Ans: In genetic engineering, Plasmid and Bacteriophage are the two common
vectors that are used.

3. Some enzymes are considered molecular scissors in genetic engineering.

What is the name assigned to such enzymes?
Ans: In genetic engineering, restriction Enzymes are the enzymes that are
responsible for the digestion of DNA strands resulting in the formation of
fragments, thus they are called molecular scissors.

4. Write conventional nomenclature of EcoRI.

Ans: E. Escherichia is a bacterium where co stands for coli; R stands for the
Name of Strain; I is the order in which the enzyme is isolated from the bacterial
strain.

5. A linear DNA fragment and a plasmid have three restriction sites for
EcoRI. How many fragments will be produced from linear DNA and plasmid
respectively?
Ans: The linear DNA will produce 4 fragments while the plasmid produces 6
fragments after their digestion by a restriction enzyme.

6. An extra-chromosomal segment of circular DNA of a bacterium is used to

carry the gene of interest into the host cell. What is the name given to it?
Ans: In bacterium, Plasmid is the extra-chromosomal segment of circular DNA
that is useful in carrying the desired gene into the host cell.

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7. Identify the recognition sites in the given sequences at which E. coli will be
cut and make sticky ends.
5 ́-GAATTC-3 ́
3 ́-CTTAAG-5 ́
Ans: 5 − G ↓ TAATTC3
3 − CTTAA ↑ G5 are the recognition sites that will make the sticky ends after
they will be cut.

8. Name the substance used as a medium in gel electrophoresis.

Ans: In the gel electrophoresis, Agarose will be a substance that will be used as
a medium.

9. What is Bioconversion?
Ans: The process of conversion of raw materials into useful products with the
help of various factors that include microbial, plant, or animal cells is called
bioconversion.

10. Name the bacterium that yields thermostable DNA polymerase.

Ans: The thermostable DNA polymerase can be produced with the help of a
bacterium which is named Thermusaquaticus.

11. Which enzymes are known as “molecular Scissors”?

Ans: The Restriction Endonuclease Enzymes are the enzymes that are
responsible for the digestion of DNA strands resulting in the formation of
fragments, thus they are called molecular scissors.

12. Name the commonly used vector for transformation in a plant cell?
Ans: The commonly used vector which is responsible for the transformation in
plants cells is named Agrobacterium tumefacient.

13. Name the technique used for amplification of DNA?

Ans: The DNA amplification can be done by the technique named the
Polymerase Chain Reaction.

14. Name the enzyme responsible for removal of 5 – phosphate group from
nucleic acid?
Ans: To remove the 5 – phosphate group from nucleic acid, Alkaline Phosphates
are the enzymes that are responsible.

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15. Who isolated Restriction enzymes for the first time?
Ans: The Restriction enzymes were first isolated by Warner Arber & Hamilton-
Smith.

16. Why do eukaryotic cells do not contain restriction enzymes?

Ans: In eukaryotic cells, the restriction enzymes are absent because the DNA is
found to be methylated heavily.

17. Why does DNA moves towards the anode in the gel electrophoresis.
Ans: In the gel electrophoresis, the DNA is found to be moving towards the anode
because the DNA is negatively charged due to the presence of a phosphate group
that results in the movement of DNA towards the anode.

Short Answer Questions 2 Marks

1. Name two main steps which are collectively referred to as the down
streaming process. Why is this process significant?
Ans: Separation and Purification are the two main steps that are together referred
to as the down streaming process.
This process is essential because the product needs to undergo clinical trial and
quality control before it reaches the market.

2. How does plasmid differ from chromosomal DNA?

Ans: The differences between plasmid and chromosomal DNA are:

Plasmid DNA Chromosomal DNA

The DNA found here is circular The DNA found here is linear.
It is found only in prokaryotic cells It is found in both prokaryotic cells
(bacterial cells). (bacterial cells) as well as
eukaryotic cells.
It is used in rDNA technology as a In rDNA technology, it is not used
vector. as a vector.

3. A bacterial cell is shown in the figure given below. Label the part ‘A’ and
‘B’. Also mention the use of part ‘A’ in rDNA technology.

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Ans: Part A is labeled as Plasmid while part B is labeled as Nucleoid
The function of a plasmid is that, in rDNA technology, it acts as a vector that
helps in transferring the desired gene into the host cell.

4. Mention two classes of restriction enzymes. Suggest their respective roles.

Ans: Exonucleases and endonucleases are two classes of restriction enzymes.
The function of Exonucleases is to remove the nucleotides from the ends of the
DNA while the Endonucleases play a major role in cutting the DNA at specific
sites between the ends of DNA.

5. In the given process of separation and isolation of DNA fragments, some

of the steps are missing, Complete the missing steps –
A: Digestion of DNA fragments using restriction endonucleases
↓
B: ..............................................................
↓
C: Staining with ethidium bromide
↓
D: Visualisation in U.V. light
↓
E: .............................................................
↓
F: Purification of DNA fragments.
Ans: At step B the process of Gel Electrophoresis takes place while at step E the
process of Elution occurs.

6. Write any two properties of restriction endonuclease enzymes?

Ans: The properties of restriction endonuclease enzymes are:
(i) The Restriction endonuclease enzymes bind at the recognition sequence of the
DNA after inspecting the length of the DNA sequence.
(ii) Its function is to cut the sugar-phosphate backbone at specific sites.

7. What are ‘Selectable markers’? What is their use in genetic engineering?

Ans: The host cells that contain the vector can be selected with the help of a gene
called a selectable marker that results in the elimination of the non–transformant.

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It is used in genetic engineering for e.g. – the gene that encodes resistance towards
the antibiotics are found to be useful selectable markers as they insert into a cell
and result in the selective growth of transformants only.

8. How can the desired product formed after genetic engineering be

produced on a commercial scale?
Ans: To make a commercial scale-out of the desired product that is formed after
the process of genetic engineering there are a series of processes that need to be
followed which are collectively called downstream processing and then it will
lead to the final processes. The final processes that are involved in the
downstream processing are Separation and purification.

9. What is “Insertional Inactivation”?

Ans: The insertional inactivation is the process of insertion of the recombinant
DNA into the coding Sequence of enzyme B– galactosidase leading to the
inactivation of the enzyme. An example is when the insert is absent in the plasmid
of bacteria then it will lead to the insertional inactivation leading to the production
of colorless colonies instead of blue-colored colonies due to the presence of
chromogenic substrate.

10. What are the two basic techniques involved in modern Biotechnology?
Ans: The two basic techniques involved in modern Biotechnology are:
a) Genetic Engineering - the technique which involves the introduction of the
genome into another host organism or results in the alternation of the nature of
genetic material that leads to change in its phenotype.
b) Techniques that are performed under sterile conditions are for the
manufacturing of a large number of the desired microbes or cells by the process
of multiplication and growth.

11. Represent diagrammatically the E. coli. Cloning vector PBR 322.

Ans: The diagram of E. coli. Cloning vector PBR 322 is represented as:

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12. Differentiate between plasmid DNA and chromosomal DNA?
Ans: The difference between plasmid DNA and chromosomal DNA are:

Plasmid DNA Chromosomal DNA

It is an extra-chromosomal part of It is nuclear or genetic DNA.
DNA.
It is found inside the protoplasm. It is found inside the nucleus.

13. What is the role of enzyme “Ligase” in genetic Engineering?

Ans: In genetic engineering, the enzyme “Ligase” acts as a molecular Suture that
helps in binding the two DNA pieces together. This process of joining the two
DNA pieces requires energy in the form of ATP and results in the formation of a
phosphodiester bond between the two cohesive ends of DNA.

14. Name the components a bioreactor must possess to achieve the desired
product?
Ans: The bioreactors are the devices in the form of the vessel which contains
various organisms or chemical substances that undergo the chemical processes
and result in the formation of the biologically active substances. A bioreactor
must consist of the following components that result in the formation of the
desired product. These components include temperature, substrate, pH, oxygen,
vitamins, and salts.

15. The following proteins of given molecular weight are Subjected to Get
electrophoresis. Write the order of Sequence in which these proteins are
isolated in a gel?

S.no. Proteins Mol. wt

1 Albumin 23,000
2 Keratin 48,000
3 Myosin 1,25,000
4 Hemoglobin 84,000
5 Ribozyme 62,000
6 Insulin 1,14,000

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Ans: The sequence of proteins obtained from top to bottom in a gel:
Myosin > Insulin >Haemoglobin> Ribozyme > Keratin > Albumin.

16. How is gene Z used as a marker?

Ans: The Lac Z Gene Is responsible for the coding of Β-galactosidase enzyme
this results in the inactivation of the enzyme due to the insertion of recombinant
DNA in the coding sequence of an enzyme Β-galactosidase. The bacterial colony
normally produces blue-colored colonies by they get the insert into their plasmid
then it will result in the production of colonies that are colorless.

17. What is Bioreactor? What are the advantages of Stirred tank Bioreactor
over Shake flask? Show diagrammatically a simple Stirred tank Bioreactor?
Ans: The bioreactors are the devices in the form of the vessel which contains
various organisms or chemical substances that undergo the chemical processes
and result in the formation of the biologically active substances. They consist of
large vessels where the raw materials using microbial, plant, animal, or human
cells are converted biologically into specific proteins. The advantages of
Bioreactor over shake flask are:
a) To produce the optimum growth of the desired product, it provides the optimal
conditions e.g., temp, pH, etc.
b) For testing the sample, a small volume of cultures can be withdrawn
periodically from the bioreactor.
c) It has an agitation system, temp control system, from control system & pH
control system.
d)

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Short Answer Questions 3 Marks

1. Since DNA is a hydrophilic molecule, it cannot pass through cell

membranes. Name and explain the technique with which the DNA is forced
into
(i) a bacterial cell
Ans: In the bacterial cell, the DNA can enter by the technique of Chemical
treatment and may be exposed to cold and high temp (42°C) alternatively.

(ii) a plant cell

Ans: The DNA in the plant cell can enter through the technique called Biolistic
or gene gun.

(iii) an animal cell.

Ans: The DNA in the animal cell can enter through the technique called Micro-
injection.

2. How will you obtain purified DNA from a cell?

Ans: Various enzymes are used to treat cells that will result in the release of DNA.
Various enzymes are cellulose (plant cells), Lysozyme (bacteria), and chitinase
(fungus) while ribonuclease and protease enzymes are used for the treatment of
and removal of RNA and proteins respectively.

3. In recombinant DNA technology, vectors are used to transfer a gene of

interest in the host cells. Mention any three features of vectors that are most
suitable for this purpose.
Ans: The vectors in the recombinant DNA technology are utilized in transferring
the gene of interest in the host cells due to the following features of vectors:
(i) Vectors constitute of origin of replication (Ori) where the gene of interest
attaches.
(ii) They constitute various selectable markers for the various genes of interest.
(iii) They are also composed of atleast one recognition site.

4. Why is Agrobacterium-mediated genetic engineering transformation in

plants considered natural genetic engineering?
Ans: Agrobacterium tumefaciens is a pathogen in many dicot plants due to its
ability to deliver a piece of DNA (TDNA) that results in the transformation of
normal plant cells into a tumor cell leading to the production of chemicals by
these tumor cells that are required by the pathogen.

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5. Observe the given sequence of nitrogenous bases on a DNA fragment and
answer the following question
5 ́ - CAGAATTCTTA - 3 ́
3 ́ - GTCTTAAGAAT - 5 ́
(a) Name a restriction enzyme that can recognize this DNA sequence.
Ans: EcoRI is the name of a restriction enzyme that helps in the recognition of
various sequences of DNA.

(b) Write the sequence after digestion.

(c) Why are the ends generated after digestion called sticky ends?
Ans: The ends that are obtained after the digestion of the DNA sequences are
called the sticky ends because they result in the formation of hydrogen bonds with
their complementary cut parts.

6. A selectable marker is used in the section of recombinants on the basis of

their ability to produce color in presence of chromogenic substrate.
(a) Mention the name of the mechanism involved.
Ans: The insertional inactivation is the process of insertion of the recombinant
DNA into the coding Sequence of enzyme B– galactosidase leading to the
inactivation of the enzyme.

(b) Which enzyme is involved in the production of color?

Ans: The b-galactosidase is the enzyme that helps in the production of color in
presence of chromogenic substrate.

(c) How is it advantageous to overuse antibiotic-resistant genes as a

selectable marker?
Ans: The use of antibiotic-resistant genes as a selectable marker is more
beneficial because due to the inactivation of antibiotics the selection of
recombinants requires simultaneous plating on two plates that are having
different antibiotics.

7. Mention the important properties which a good vector must possess?

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Ans: The important properties which a good vector must possess are -
i) Size - The size of the vector must be small so that it helps them in purification
and isolation easily.
ii) Origin of replication – A site where replication starts and is made up of the
sequence of base pairs. When the DNA is attached to this sequence then it will
result in the replication within its host cell & thus, controls the number of
copies of linked DNA.
iii) Selectable Marker - The host cells that contain the vector can be selected with
the help of a gene called a selectable marker that results in the elimination of
the non–transformant. It is used in genetic engineering.
iv) Cloning Sites – The site in the vector where the foreign/alien DNA attaches is
also called the unique recognition site. A particular restriction enzyme will cut
the vector-only at a particular recognition site.

8. Describe any three vectors less method of introducing the rDNA into a
competent host cell?
Ans: The three vectors less method of introducing the rDNA into a competent
host cell are:
i) Transformation: The bacterial cell is first treated with the specific concentration
of divalent cation e.g., Ca2+ so that they can be competent enough to take up
the DNA in the plasmid. The calcium ions increase the efficiency of DNA to
enter into a bacterium through pores in its cell wall. By the process of
incubating the cells with recombinant DNA on ice, then placing them at 420
C, and then again putting them back into ice the recombinant DNA can then be
forced into cells. This helps the bacteria in taking up the recombinant DNA.
ii) Microinjection: With the help of a microneedle of the tip with a diameter (~
4mm), the recombinant DNA can be injected directly into the nucleus of an
animal cell.
iii) Biolistic / Gene gun: the cells of the DNA that are coated with particles of
gold or tungsten are bombarded with high-velocity micro-gun.

9. Why is Agrobacterium-mediated genetic transformation described as

Natural Genetic engineering in plants?
Ans: The Agrobacterium tumefaciens-interceded plant genetic transformation
measure requires the presence of two genetic segments situated on the bacterial
Ti-plasmid. Basically, the main basic part is the T-DNA, which is characterized
by conserved 25-base pair imperfect repeats at the closures of the T-region known
as a border sequence. The tumor-inducing (Ti) plasmid is used as a cloning vector
to transfer the desired gene into plants as they insert a part of their DNA in plants
during the infection. The second is the virulence (vir) region, which is made out

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of in any event seven significant loci (virA, virB, virC, virD, virE, virF, and virG)
encoding parts of the bacterial protein machinery which is mediating the T-DNA
processing and transfer. The gene of interest is attached to the T-DNA so that it
automatically gets transformed into plant cells thus, Agrobacterium tumefacien
is known as the “Natural Genetic Engineer” of plants.

10. Mention the important tools required for genetic engineering

technology?
Ans: The process of genetic engineering is accomplished only when we have the
following key tools:
a) Restriction enzymes: In genetic engineering, restriction Enzymes are the
enzymes that are responsible for the digestion of DNA strands resulting in the
formation of fragments, thus they are called molecular scissors. They are of
types: Endonucleases and Exonucleases.
b) Cloning Vector: The cloning Vector is the part of the DNA molecule that is
attached to the DNA Segment of an organism that is desired and then transfers
into the cell or DNA of another organism.
c) Desired foreign DNA: The desired foreign DNA segment is the part of DNA
that consists of genes having desired characters that are being transferred into
the genome of another cell with the help of the cloning vector.

Long Answer Questions 5 Marks

1. The development of bioreactors is required to produce large quantities of

products.
(a) Give optimum growth conditions used in bioreactors.
Ans: The bioreactors are the devices in the form of the vessel which contains
various organisms or chemical substances that undergo the chemical processes
and result in the formation of the biologically active substances. To produce the
optimum growth of the desired product the optimum temperature, pH, substrates,
salts, vitamins, and oxygen are required.

(b) Draw a well-labeled diagram of a simply stirred tank bioreactor.

Ans: A simply stirred tank bioreactor is shown below:

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(c) How does a simply stirred tank bioreactor differ from sparged stirred –
tank’ bioreactor?
Ans: In the simply stirred tank bioreactor the stirrer facilitates the even mixing
and the oxygen availability throughout the process, whereas for proper mixing
throughout the reactor in the case of sparged stirred-tank bioreactor the air is
found to be bubbled.

2. In the given figure, one cycle of polymerase chain reaction (PCR) is shown-

(a) Name the steps A, B, and C.

Ans: A: Denaturation, B: Annealing, and C: Extension.

(b) Give the purpose of each of these steps.

Ans: The purpose of each step is:

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(A) Denaturation: Due to high temperature, the heat will break or denature the
two complementary strands of DNA and results in their separation.
(B) Annealing: The hybridization of the denatured DNA strands takes place with
the help of the primers.
(C) Extension: The target DNA sequence will synthesize its copies by the process
of the extension of the primers.

(c) State the contribution of the bacterium Thermus Aquaticus in this

process.
Ans: The Taq polymerase enzyme is found to be isolated from the bacterium
Thermus Aquaticus which functions at a very high temperature and results in the
denaturation of double-stranded DNA.

3. Study the figure of vector pBR322 given below in which foreign DNA is
ligated at the Bam H1 site of the tetracycline resistance gene.

Answer the following questions:

(a) Mention the function of rop.
Ans: The rop is responsible for the coding of the proteins that are involved in the
replication of plasmids.

(b) What will be the selectable marker for this recombinant plasmid and
why?
Ans: The selectable marker for this recombinant plasmid will be the ampicillin
resistance gene. They after placing them on an ampicillin-containing medium will
undergo the process of plating that will help in the differentiation between the
trAns.formants from non-trAns.formants.

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(c) Explain transformation.
Ans: Transformation is the process of transferring DNA from one cell and then
placing them into the other cell which will result in the formation of the
recombinant cell consisting of the properties of both the cells.

4. Describe the various steps involved in Recombinant DNA technology with

the help of a well labeled. Diagram?
Ans: The steps involved in the recombinant DNA are:
i) Identification of DNA with desirable Genes: The addition of chilled ethanol
will result in obtaining the purified DNA while by using various other
appropriate techniques the other molecules in the target cell can also be
removed.
ii) Cutting the DNA at a specific location: The source and vector DNAS after
being cut will be introduced together having a gene of interest and specific
restriction site and will be joined together with the help of an enzyme called
ligase.
iii) Insertion of Recombinant DNA into host cell: The bacterial cell is first treated
with the specific concentration of divalent cation e.g., Ca2+ so that they can be
competent enough to take up the DNA in the plasmid. The calcium ions
increase the efficiency of DNA to enter into a bacterium through pores in its
cell wall. It is a vector-less method.

iv) Selection & Screening: The selectable marker for this recombinant plasmid
will be the ampicillin resistance gene. They after placing them on an

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 ampicillin-containing medium will undergo the process of plating that will
help in the differentiation between the trAns.formants from non-
trAns.formants.
v) Obtaining the foreign Gene product: The gene of interest after its cloning along
with the presence of optimum growth conditions will result in the expression
of target proteins which then needs to be produced on a large scale.

5. Expand PCR? Describe the different steps involved in this technique?

Ans: The Polymerase Chain Reaction is the method of making millions of DNA
copies from a DNA sample. It has two main reagents: primers (short single-
stranded DNA fragments that are a complementary sequence to the target DNA),
and DNA polymerase. The DNA polymerase is heat stable, that is Taq
polymerase which is extracted from the bacteria Thermus aquaticus. Each cycle
has three steps:
a) DENATURATION: In the first step, the two strands of the DNA helix are
physically separated at a heat during a process called macromolecule
denaturation.
b) RENATURATION / ANNEALING: In the second step, the temperature is
lowered so that the primers can bind to the complementary sequences of DNA.
c) EXTENSION: The third step is the target DNA sequence will synthesize its
copies by the process of the extension of the primers. The temperature is raised
to 750c. At this temperature, Taq – polymerase initiates DNA Synthesis at the
3-OH end of the primer.

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6. What are Restriction enzymes? Why do bacteria have these restriction
enzymes? Show diagrammatically a restriction enzyme its recognition & the
product it produces?
Ans: Restriction Enzymes are the endonuclease enzymes that are responsible for
the digestion of DNA strands resulting in the formation of fragments, thus they
are called molecular scissors. They are found in bacteria cells as they help in
cutting the foreign DNA and results in the modification of the restriction system
and thus results in the improving of the immunity in the bacterial cell.
Name of Restriction enzyme- EcoRI Substrate DNA on which it acts

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