ADITYA VIDYASHRAM RESIDENTIAL SCHOOL
CLASS XII (2024-2025)
WORK SHEET NO. 14
CHAPTER -9 – BIOTECHNOLOGY PRINCIPLES & PROCESSES
1. Biotechnologists refer to Agrobacterium
tumifaciens as a natural genetic engineer of plants. 11. How is Agrobacterium tumifaciens able to
Give reasons to support the statement. transform a normal plant cell into a tumor?
OR
Why is Agrobacterium tumefaciens a good cloning 12. How can retroviruses be used efficiently in
vector? biotechnology experiments inspite of them being
disease causing?
2. Suggest a technique to a researcher who needs to
separate fragments of DNA. 13. State what happens when an alien gene is
ligated at PvuI site of PBR322 plasmid.
3. List the key tools used in recombinant DNA
technology. 14. Why is 'Plasmid' an important tool in
biotechnology experiments?
4. What is EcoRI? How does EcoRI differ from an
exonuclease? 15. Name the specific sequence of DNA in a
plasmid that the 'gene of interest' ligates with, to
5. How can bacterial DNA be released from the enable it to replicate.
bacterial cell for biotechnology experiments.
16. State the role of DNA ligase in biotechnology.
6. Write the two components of the first artificial
recombinant DNA molecule constructed by Cohen 17. Mention the use of cloning vector in
and Boyer. biotechnology.
7. Mention the type of host cells suitable for the 18. Why is it essential to have a selectable marker
gene guns to introduce in alien DNA. in a cloning vector?
8. Name the host cells in which microinjection 19. Why do DNA fragments move towards the
technique is used to introduce an alien DNA. anode during gel-electrophoresis?
9. Name the material used as matrix in gel
electrophoresis and mention its role. 20. In the year 1963, two enzymes responsible for
restricting the growth of bacteriophage in E. coli
10. State what happens when an alien gene is were isolated. How did the enzymes act to restrict
ligated at salI site of pBR322 plasmid. the growth of the bacteriophage?
WORK SHEET NO. 15
21. How is the action of exonuclease different from 24. What is the role of ethidium bromide during
that of endonuclease? agarose-gel electrophoresis of DNA fragments?
22. Mention the role of molecular scissors in 25. Write the importance of the bacterium Thermus
recombinant DNA technology. aquaticus in polymerase chain reaction.
23. Name the technique used for separating DNA 26. Mention the source of thermostable DNA
fragments in the laboratory. polymerase.
27. Why is it not possible for an alien DNA to 36. Name the type of bioreactor shown. Write the
become part of a chromosome anywhere along its purpose for which it is used.
length and replicate normally?
28. What is the host called that produces a foreign
gene product? What is this product called?
29. Why is the enzyme cellulase used for isolating
genetic material from plant cells but not for animal
37. State how has Agrobacterium tumifaciens been
cells?
made a useful cloning vector to transfer DNA to
plant cells.
30. What is a plasmid?
38. How are sticky ends' formed on a DNA strand?
31. What are Cry genes? In which organism are
Why are they so called?
they present?
39. A schematic representation of Polymerase
32. Discuss the role the enzyme DNA ligase plays
Chain Reaction (PCR) up to the extension stage is
during DNA replication.
given below. Answer the questions that follow:
33. Name the source organism that possesses Taq
polymerase. What is so special about the function of
this enzyme?
OR
Why is Taq polymerase preferred in PCR? Mention
the source of this enzyme.
OR
Explain the contribution of Thermus aquaticus in
the amplification of a gene of interest.
34. Explain the work carried out by Cohen and
Boyer that contributed immensely in biotechnology.
(i) Name the process A. (ii) Identify B. (iii) Identify
C and mention its importance in PCR.
35. Any recombinant DNA with a desired gene is
required in billion copies for commercial use. How
40. Write the role of ori and restriction site in a
is the amplification done? Explain
cloning vector pBR322.
WORK SHEET NO. 16
41. How does a restriction nuclease function? 45. (a) Mention the difference in the mode of action
Explain. of exonuclease and endonuclease. (b) How does
42. How is insertional inactivation of an enzyme restriction endonuclease function?
used as a selectable marker to differentiate
recombinants from nonrecombinants? 46. (a) Explain how to find whether an E. Coli
43. Explain palindromic nucleotide sequence with bacterium has transformed or not when a
the help of a suitable example. recombinant DNA bearing ampicillin-resistant gene
44. Why is making cells competent essential for is transferred into it. (b) What does the ampicillin-
biotechnology experiments? List any two ways by resistant gene act as in the above case?
which this can be achieved. ] 47. Why and how bacteria can be made
'competent'?
48. Write any four ways used to introduce a desired 56. A and B are the two different cloning vectors in
DNA segment into a bacterial cell in recombinant two different bacterial colonies cultured in
technology experiments. chromogenic substrate. Bacterial colonies with
49. How is the bacterial cell made competent to coloning vector A were colourless whereas those
receive recombinant DNA? Explain. with B were blue coloured. Explain giving reasons
the cause of the difference in colour that appeared
50. (a) How is an exonuclease functionally different 57. How is DNA isolated in purified form from a
from an endonuclease? (b) Give an example of any bacterial cell?
two endonucleases other than SalI. 58. Name the source organism from which Ti
51. State the role of UV-light and ethidium bromide plasmid is isolated. Explain the use of this plasmid
during gel electrophoresis of DNA fragments. in biotechnology.
52. Explain the role of Ti plasmids in 59. Name the natural source of agarose. Mention
biotechnology. one role of agarose in biotechnology.
53. How are recombinant vectors created? Why is 60. Study the linking of DNA fragments shown
only one type of restriction endonuclease required above. (a) Name 'A' DNA and 'B' DNA. (b) Name
for creating one recombinant vector? the restriction enzyme that recognises this
54. A recombinant DNA is formed when sticky palindrome. (c) Name the enzyme that can link
ends of vector DNA and foreign DNA join. Explain these two DNA fragments.
how the sticky ends are formed and get joined.
55. (a) Illustrate the recognition sequence of EcoRI
and mention what such sequences are called? (b)
How does restriction endonuclease act on a DNA
molecule?
WORK SHEET NO. 17
61. How do 'ori' and 'cloning sites' facilitate cloning 68. Explain the action of the restriction
into a vector? endonuclease EcoRI.
62. Name two commonly used bioreactors. State the 69. How are the DNA fragments separated by gel
importance of using a bioreactor. electrophoresis visualised and separated for use in
constructing recombinant DNA?
63. Name the source of the DNA polymerase used
in PCR technique. Mention why it is used. 70. Name and explain the technique used for
separating DNA fragments and making them
64. (a) Mention the number of primers required in available for biotechnology experiments.
each cycle of polymerase chain reaction (PCR).
Write the role of primers and DNA polymerase in 71. What are cloning sites in a cloning vector?
PCR. (b) Give the characteristic feature and source Explain their role. Name any two such sites in
organism of the DNA polymerase in PCR. pBR322.
65. What is genetic engineering? List the steps in 72. How are the DNA fragments separated and
rDNA technology. isolated for DNA fingerprinting? Explain.
66. Name the organism from where the 73. How are the following used in biotechnology?
thermostable DNA polymerase is isolated. State its (a) Plasmid DNA (b) Recognition sequence (c) Gel
role in genetic engineering. electrophoresis
67. Explain with the help of a suitable example the
naming of a restriction endonuclease. 74. How is the amplification of a gene sample of
interest carried out using Polymerase Chain
Reaction (PCR)?
OR
Explain in sequence the process of amplification of
a gene of interest using polymerase chain reaction.
OR WORK SHEET NO. 18
How is repeated amplification of DNA made 81. Rearrange the following in the correct sequence
possible during PCR? to accomplish an important biotechnological
reaction: (a) In vitro synthesis of copies of DNA of
75. (a) Identify (A) and (B) illustrations in the interest (b) Chemically synthesized oligonucleotides
following (c) Enzyme DNA-polymerase (d) Complementary
region of DNA (e) Genomic DNA template (f)
Nucleotides provided (g) Primers (h) Thermostable
DNA-polymerase (from Thermus aquaticus)
82.
(b) Write the term given to (A) and (C) and why?
(c) Expand PCR. Mention its importance in
biotechnology
76. Prepare a flow chart in formation of
recombinant DNA by the action of restriction
endonuclease enzyme EcoRI.
OR
How does a restriction endonuclease function?
Name a restriction endonuclease and write the
specific sequence of bases that it recognizes. (a) Identify the selectable markers in the diagram of
E. coli vector shown above. (b) How is the coding
77. How can DNA segments, separated by gel sequence of b– galactosidase considered a better
electrophoresis, be visualised and isolated? marker than the ones identified by you in the
diagram? Explain.
78. (a) Why must a cell be made ‘competent’ in OR
biotechnology experiments? How does calcium ion (a) A recombinant vector with a gene of interest
help in doing so? (b) State the role of ‘biolistic gun’ inserted within the gene of b-galactosidase enzyme,
in biotechnology experiments. is introduced into a bacterium. Explain the method
that would help in selection of recombinant colonies
79. (a) Name the selectable markers in the cloning from non-recombinant ones. (b) Why is this method
vector pBR322? Mention the role they play. (b) of selection referred to as “insertional inactivation”?
Why is the coding sequence of an enzyme (b-
galactosidase) a preferred selectable marker in 83. Draw a schematic sketch of pBR 322 plasmid
comparison to the ones named above? and label the following in it: (a) Any two restriction
OR sites. (b) Ori and rop genes. (c) An antibiotic
Why does the ‘insertional inactivation’ method to resistant gene.
detect recombinant DNA is preferred to ‘antibiotic
resistance’ procedure? 84. Many copies of a specific gene of interest are
required to study the detailed sequencing of bases in
80. Explain the role of the enzyme EcoRI in it. Name and explain the process that can help in
recombinant DNA technology. developing large number of copies of this gene of
OR interest.
Explain with the help of an example the relationship OR
between restriction endonuclease and a palindromic Suggest and describe a technique to obtain multiple
nucleotide sequence. copies of a gene of interest in vitro.
85. Draw a labelled sketch of sparged-stirredtank formed on both the segments where the two DNA
bioreactor. Write its application. segments will join later to form a recombinant
DNA.
86. State the functions of the following in the
cloning vector pBR322: (a) Ori (b) rop, and (c) 96.
HindIII sites
87. Draw pBR322 cloning vector, Label 'ori', 'rop'
and any one antibiotic resistance site on it and state
their functions.
88. Draw a schematic diagram of the E. coli cloning
vector pBR322 and mark the following in it: (a) ori
(b) rop (c) ampicillin resistance gene (d)
tetracycline resistance gene (e) restriction site
BamHI (f) restriction site EcoRI (a) Name the organism in which the vector shown
89. (a) Draw schematic diagrams of segments of a is inserted to get the copies of the desired gene. (b)
vector and a foreign DNA with the sequence of Mention the area labelled in the vector responsible
nucleotides recognised by EcoRI. (b) Draw the for controlling the copy number of the inserted
vector DNA segment and foreign DNA segments gene. (c) Name and explain the role of a selectable
after the action of EcoRI and label the sticky end marker in the vector shown.
produced.
97. (a) EcoRI is a restriction endonuclease. How is
90. (a) Explain the basis on which the gel it named so? Explain. (b) Write the sequence of
electrophoresis technique works. (b) Write any two DNA bases that the enzyme recognises. Mention the
ways the products obtained through this technique point at which the enzyme makes a cut in the DNA
can be utilised. segment.
91. Explain the importance of – (a) Restriction 98. (a) Name the technique used for separation of
Endonuclease, (b) "Ori" and (c) gel-electrophoresis DNA fragments. (b) Write the type of matrix used
in recombinant DNA technology. in this technique. (c) How is separated DNA
visualised and extracted for use in recombinant
92. Why has a bacterium to first become competent technology?
to be able to take up DNA? Explain how it becomes
competent and takes in the recombinant DNA 99. Name and explain the technique used in the
separation and isolation of DNA fragments to be
93. How is the desired DNA for biotechnology used in recombinant DNA technology.
experiments first fragmented and later separated by
gel-electrophoresis? Explain. 100. (a) What is EcoRI? What does 'R' represent in
this? (b) Give the palindromic nucleotide sequence
94. (a) Why are restriction endonucleases so called? recognised by it. (c) Explain its action.
(b) What is a palindromic nucleotide sequence?
How do restriction endonucleases act on WORK SHEET NO. 19
palindromic sites to create 'sticky ends'?
101. Why are genes encoding resistance to
95. EcoRI is used to cut a segment of foreign DNA antibiotics considered useful selectable markers for
and that of a vector DNA to form a recombinant E. coli cloning vector? Explain with the help of one
DNA. Show with the help of schematic diagrams. example.
(a) The set of palindromic nucleotide sequence of
base pairs the EcoRI will recognise in both the 102. (a) What are 'molecular scissors'? Give one
DNA segments. Mark the site at which EcoRI will example. (b) Explain their role in recombinant DNA
act and cut both the segments. (b) Sticky ends are technology.
103. Why is Agrobacterium tumefaciens a good
cloning vector? Explain.
104. DNA being hydrophilic cannot pass through
the cell membranes of a host cell. Explain how does
recombinant DNA get introduced into the host cell
to transform the latter.
114. a) Identify the enzymes A, B and C.
105. Explain any three methods to force 'alien' or b) Plasmid was used as a vector in this
recombinant DNA into host cells. investigation. Name another vector that can be used
here.
106. (a) Explain the significance of 'palindromic When two different varieties– a conventional
nucleotide sequence' in the formation of variety and a GM crop of corn crop were grown in a
recombinant DNA. (b) Write the use of restriction field, it was noticed that corn borers attacked only
endonuclease in the above process. the conventional variety.
a) Suggest a suitable treatment using genetic
107. Describe the roles of heat, primers and the engineering approach for damage control in the
bacterium Thermus aquaticus in the process of conventional variety. Justify your approach.
PCR. b) Name the gene associated for development of
GM crop for the control of this pest. Explain its
108. (a) How has the development of bioreactor impact on the insect pest?
helped in biotechnology? (b) Name the most c) How does it not harm the source from which it is
commonly used bioreactor and describe its working. taken?
109. Explain the roles of the following with the help OR
of an example each in recombinant DNA Factor VIII protein is a very useful protein for blood
technology: (a) Restriction Enzymes (b) Plasmids clotting in the human body. If deficient, it can either
be plasma derived or can be made as a genetically
110. (i) Describe the characteristics a cloning vector engineered recombinant protein.
must possess. (ii) Why DNA cannot pass through
the cell membrane? Explain. How is a bacterial cell a) Name a genetic disease that may be treated using
made ‘competent’ to take up recombinant DNA recombinant human factor VIII.
from the medium?
b) Before recombinant human factor VIII was
111. If a desired gene is identified in an organism available, this disease was treated with factor VIII
for some experiments, explain the process of the received from donated blood. Give two possible
following: (a) Cutting this desired gene at specific advantages of using recombinant human factor VIII
location (b) Synthesis of multiple copies of this instead of it being obtained from donated blood, to
desired gene treat this disease.
112. (a) Describe the different steps in one complete c) What is unique feature in inheritance pattern of
cycle of PCR. (b) State the purpose of such an the disease as mentioned in part (a) above.
amplified DNA sequence.
115. Given below is an image showing a special
113. In an effort to mass produce a useful protein, situation in which a dsRNA from a source has been
scientists extracted a precise section of mRNA from introduced into a host cell.
the cytoplasm of cells which naturally produced the
protein. The mRNA strand was then used as a
template to enzymatically synthesise a strand of
complementary DNA (cDNA).
for the control of this pest.
b) What effect will the spores of Bt have
on the insect pest?
c) How has field B developed resistance
against this pest?
OR
Lipoprotein lipase deficiency (LPLD)is a
genetic disorder in which a person has a
defective gene for lipase. This leads to
high triglycerides, stomach pain, fat
a) How can dsRNA of nematode specific gene be deposits under the skin. It may eventually
introduced into the host cell as shown in the figure affect the liver, pancreas and may also
given above? cause diabetes. The disorder occurs if a
b) What can be the source of dsRNA for this child acquires defective genes from both
process?
c) What will be the impact of this interaction 118. The structure below shows pUC18 which is
between dsRNA and mRNA on the mRNA and similar to pBR322 in its function. However, they
entire cellular machinery of host plant? differ in some of their restriction sites and number
of ori. The ori number for pBR322 is approximately
116. The image below shows the result of plating 20.
bacteria in chromogenic medium after incorporating
the gene of interest in plasmid. Some plates had
blue colonies; some plates had white colonies. A
single bacterium extracted from Plate I,II,III is
shown below:
a) How are puc18 and pBR322 used in
biotechnological studies? OR What will be the
impact if oriin the above structure gets damaged? b)
The lac z gene has many recognition sites. Study the
segment of DNA given below and answer the
questions 5’... ATC GTA AAG CTT CAT…3’ 3’...
TAG CAT TTC GAA GTA…5’ i) Applying your
On the basis of your observations a) Identify the knowledge of palindrome sequences identify and
plate(s) which is/are white. Give a reason. b) mark the possible region where the restriction
Identify the plate(s) which is/are blue. Give a enzyme X will act. ii) Restriction enzyme Y was
reason. used to extract gene of interest from a plant. This
gene needs to be inserted in the given DNA
117. segment which has been treated with restriction
A farmer grew 2 varieties of corn crop in enzyme X. Will there be a successful
field A and B. He grew normal corn crops recombination? Explain with a reason.
in field A and GM corn crops in field B. c) Which one of the two (pUC18 and pBR322)
He observed corn borers attacked only in would you prefer for biotechnological studies?
field A. To control it, spores of Bt were Justify.
sprayed in field A.
a) Name the gene in the spores responsible 119. Rati wants to grow a variant of the
lactobacillus spps. in a bioreactor. Lactobacillus is
an anaerobic bacterium commonly used as a starter If the vector was cut using HindIII, what would
culture for diary products. Shown colonies growing on a medium containing
below is a bioreactor she had in her laboratory. tetracycline DEFINITELY contain?
(a) Identify ONE component that should definitely (b) Suggest which of the three restriction enzymes
NOT be present in the reactor to would be suitable for insertion of the gene of
grow the lactobacillus spps. Justify. interest. Give TWO advantages of using the enzyme
(b) Explain TWO quantities that the sensors in the chosen by you.
bioreactor should monitor. (c) State ONE disadvantage of using the other two
restriction enzymes not chosen in (b).
120. Shown below is a cloning vector 'Z' that Kamla
wants to use to create a recombinant vector with her
gene of interest.
The vector consists of sites for three restriction
enzymes - ScaI, HindIII and EcoRI. Restriction
sites for the same enzymes are also present in the
gene of interest. There are two 'ori' sequences - one
allows it to replicate in Escherichia coli and another
allows replication in Zymomonas mobilis. Apart
from this, the vector consists of two antibiotic
resistance genes - one against tetracycline (tet) and
another against chloramphenicol (cml).
(a) What is the advantage of having two 'ori'
sequences in the same vector?
Give a situation in which this would be particularly
useful.
OR