BIOTECHNOLOGY

1) What is biotechnology according to EFB?

2) Expand EFB.
3) Which organisation has given a definition for biotechnology encompassing both traditional as well the
modern molecular view?
4) Define biotechnology.
5) Give some examples of some processes of biotechnology.
6) Can making curd, bread or wine be considered a form of biotechnology? Why can’t they be thought as
modern biotechnology?
7) How is the word biotechnology used in a restricted sense today?
8) Name any 2 core techniques that gave rise to modern biotechnology.
9) Define (a) Genetic engineering (b) Bioprocess engineering
10) Which type of reproduction permits variation and which does preserve genetic setup?
11) Name any one disadvantage of traditional breeding techniques. Suggest a solution.
12) What is meant by maintenance of sterile ambience in chemical engineering processes in
biotechnology?
13) What are some examples of products that can be manufactured through these biotechnological
processes?
14) What would happen if any alien DNA is introduced into a host as such?
15) Which sequence of chromosome initiates replication?
16) Define cloning on context of basic DNA transfer.
17) What should be done to make a foreign DNA replicate in host organism?
18) What is the first instance of the construction of an artificial recombinant DNA molecule?
19) How did Stanley Cohen and Herbert Boyer isolate the antibiotic resistance gene?
20) What is a plasmid?
21) Name the bacteria used in creation of recombinant DNA for the first time?
22) ____________ are also known as molecular scissors.
23) How do plasmid DNA act as vectors?
24) The linking of antibiotic resistance gene with the plasmid vector became possible with the enzyme
DNA ligase____________.
25) What are the 3 steps in modifying an organism genetically? Explain with antibiotic resistance cloning in
E.coli briefly.
26) What is the role of plasmid DNA in genetic engineering?
27) How does a mosquito relate to a plasmid in terms of genetic engineering?
28) What enzyme makes the linking of antibiotic resistance gene with the plasmid vector possible?
29) What is recombinant DNA?
30) Name some tools used in biotechnology for creation of r-DNA.
31) In 1963, two enzymes taking part for restriction of growth of bacteriophage were discovered. What are
these enzymes? Name their functions.
32) Name the 1st restriction endonuclease to be discovered.
33) Currently, there are around ____ number of restriction enzyme obtained from over ____ number of
strains.
34) What is the nomenclature of naming restriction endonuclease enzymes? Give example and explain.
35) Assertion – A plasmid can be used as vector to deliver an alien piece of DNA into the host organism.
Reason – Mosquito acts as an insect vector to transfer the malarial parasite into human body.
36) In EcoRI, the letter ‘R’ is derived from the name of _________. Roman numbers following the names
indicate __________________.
37) Restriction enzymes belong to a class of enzymes named __________.
38) Differentiate between exonuclease and endonuclease.
39) What is a recognition sequence in terms of restriction enzymes?
40) What are palindromes generally? What is palindromic DNA sequence? Give an example for each.
41) The restriction endonucleases cut DNA at specific points in ___________ of DNA and breaks
__________ type of bonds.
42) How is a palindrome in DNA different from a word-palindrome?
43) Explain with a neat diagram the functioning of restriction endonuclease.
44) Define sticky ends. How are they useful in forming r-DNA?
45) Assertion - Overhanging stretches called sticky ends are present on each strand after the activity of
endonuclease.
Reason – These are named so because they form hydrogen bonds with their complementary cut
counterparts.
46) Which property of sticky ends is exploited by DNA ligase?
47) Where do restriction enzymes cut the DNA?
48) What is the role of restriction endonucleases in genetic engineering?
49) Unless one cuts the vector and the source DNA with the same restriction enzyme, the recombinant
vector molecule cannot be created. Why?
50) Name the technique that can be used to separate DNA fragments.
51) Where are the fragments loaded in the separation apparatus (gel electrophoresis)?
52) Explain the working of gel electrophoresis.
53) What about the size of fragments? Do they have any effect on the distance moved?
54) The separated fragments are stained with ______________ and exposed to UV.
55) After staining, the fragments of DNA appear as ____________ coloured bands.
56) Define elution.
57) Name the medium used in gel electrophoresis.
58) Briefly explain the process of separation and isolation of DNA fragments.
59) Why can DNA fragments be separated using an electric field?
60) Where is the medium used for electrophoresis obtained?
61) How can you visualise the separated DNA fragments?
62) Name and explain some factors essential for cloning a vector.
63) What is the ability of plasmids and bacteriophages within bacterial cells?
64) How many copies of their genome do bacteriophages typically have within bacterial cells?
65) How many copies per cell do some plasmids have?
66) What happens if we link an alien piece of DNA with bacteriophage or plasmid DNA?
67) Define origin of replication.
68) What are selectable markers?
69) Define transformation.
70) What is the purpose of linking a piece of DNA to the ori sequence?
71) Define insertional inactivation.
72) Name any two selectable markers.
73) Why is the traditional selectable marker a cumbersome procedure? How does a modern one
overcome this process?
74) Explain both the methods for finding recombinants and non-recombinants.
75) What happens if there are more than one recognition sites within the vector?
76) Where is the ligation of alien DNA carried out?
77) What is an example of a site where you can ligate foreign DNA?
78) What happens when there is a presence of a chromogenic substrate?
79) How does a recombinant DNA affect the β-galactosidase enzyme?
80) What is the function of T-DNA in Agrobacterium tumifaciens?
81) How do retroviruses in animals work similarly to Agrobacterium tumifaciens in plants?
82) How has understanding gene delivery by pathogens been useful for humans?
83) How to transform a host into a competent one? What is the need to do this?
84) Explain micro-injection.
85) Explain biolsitics.
86) Biolistics is also called as ___________________
87) Why can't DNA pass through cell membranes?
88) Explain various enzymes and processes for isolation of DNA.
89) How can RNA and proteins be removed from the mixture?
90) What substance is added to make DNA precipitate out? How do you know that DNA has precipitated?
91) What is used to check the progression of a restriction enzyme digestion?
92) Expand PCR.
93) What are primers used in PCR?
94) The DNA polymerase used in PCR is obtained from the bacterium ____________________.
95) How does a PCR generate so many copies of r-DNA? Explain with a neat labelled diagram.
96) Name the 3 processes in PCR cycle.
97) What happens when a recombinant DNA bearing gene for resistance to an antibiotic is transferred into
E. coli cells?
98) What happens when the transformed cells are spread on agar plates containing ampicillin?
99) How can one select a transformed cell in the presence of ampicillin?
100) What happens when you insert a piece of alien DNA into a cloning vector and transfer it into a
bacterial, plant or animal cell?
101) What is the ultimate aim of almost all recombinant technologies?
102) Why is there a need for the recombinant DNA to be expressed?
103) What is the process called when a protein encoding gene is expressed in a heterologous host?
104) Why might there be a need for large-scale production of a target protein?
105) How can cells harbouring cloned genes of interest be grown? What can the cultures of cells
harbouring cloned genes be used for?
106) What is a continuous culture system?
107) Why can't small volume cultures yield appreciable quantities of products?
108) What are bioreactors and why were they developed?
109) How much volume of materials could be synthesised from a bioreactor?
110) Define downstream processing.
111) What must be the essential precautions taken while in case of downstream processing?
112) What made modern biotechnology using genetically modified organisms possible?
113) What is the key process in genetic engineering called?
114) What is the most common type of bioreactors?
115) Draw a neat labelled diagram of a stirring type bioreactor and sparged bioreactor.
116) What is the stage called after the biosynthetic stage in product development?
117) What are the processes involved in downstream processing?
118) What needs to be added to the product during formulation?
119) What are the 3 critical areas of research in biotechnology?
120) What are the applications of biotechnology?
121) What are the 3 options that can be used to increase the production of food?
122) Name any 2 disadvantages of green revolution and agro-based farming. Suggest a solution to
overcome these problems.
123) Expand GMO. Define GMO.
124) Give a few advantages of GM crops.
125) Give an example of biofortification using GM crops.
126) Expand Bt.
127) What is Bt toxin and what is it used for?
128) What factors contributed to increased yields during the Green Revolution?
129) Why are agrochemicals often not a viable solution for farmers in the developing world?
130) Give some examples of Bt enabled crops.
131) How has genetic modification reduced reliance on chemical pesticides?
132) How does genetic modification increase the efficiency of mineral usage by plants?
133) In what way does genetic modification help to reduce post-harvest losses?
134) Bt toxin has been produced by ________________ bacteria.
135) How is GM used for in industries?
136) How does Bt toxin work in terms of agriculture?
137) Bt toxin is toxic and hazardous to some organisms. But the bacteria producing it doesn’t die. Why?
138) What types of insects do some strains of Bacillus thuringiensis kill?
139) Name a lepidopteran, coleopteran, dipteran killed by Bt toxin.
140) What does B. thuringiensis form during a particular phase of their growth? What is their
composition?
141) How is the inactive toxin converted into an active form?
142) What are Bt toxin genes?
143) What does the choice of Bt toxin genes depend on? What is the name of the gene that codes for the
toxin?
144) Give examples of proteins encoded by cry genes and what they control.
145) Name a nematode that infects the root of tobacco plants.
146) What strategy was adopted to prevent the infestation of Meloidegyne incognitia in tobacco plants?
147) What is RNA interference (RNAi)?
148) Where could the source of the complementary RNA come from in case of RNAi?
149) How does RNAi help in defending against nematodes?
150) What is the contribution of biotechnology towards agriculture? Name any 2 of them and explain.
151) What vectors were used to introduce nematode-specific genes into the host plant?
152) ______ number of recombinant therapeutics have been in use in the world. How much of them has
been approved in India?
153) What is a significant advantage of recombinant therapeutics over similar products isolated from non-
human sources?
154) Write a short note on genetically engineered insulin.
155) What could a diabetic patient do if enough human-insulin was not available?
156) Would the insulin isolated from other animals be as effective as that secreted by the human body?
157) Why insulin can’t be taken as a drug?
158) Draw a neat structure of insulin in both inactivated as well as in activated form.
159) Where was insulin for diabetes earlier extracted from?
160) How is insulin synthesized in mammals?
161) What was the main challenge for production of insulin using rDNA techniques?
162) Which company prepared two DNA sequences corresponding to A and B chains of human insulin in
1983?
163) What is gene therapy? Give one example.
164) How does gene therapy work?
165) When was the 1st gene therapy done?
166) Name some clinical management techniques to treat ADA deficiency. Why is gene therapy preferred
over these ways?
167) How is ADA deficiency clinically treated using gene therapy?
168) What is adenosine deaminase (ADA) crucial for? What causes ADA deficiency?
169) What is a potential permanent cure for ADA deficiency?
170) Why does a patient require periodic infusion of genetically engineered lymphocytes in gene therapy?
171) Write a short note on gene therapy.
172) Write a short note on the role of biotechnology in molecular diagnostics.
173) What is the importance of early diagnosis and understanding the pathophysiology of a disease?
174) Why is early detection not possible with conventional methods of diagnosis like serum and urine
analysis?
175) What are some techniques that can serve the purpose of early diagnosis?
176) How does PCR help in detecting diseases at an early stage?
177) The principle in which ELISA works is _______________________.
178) Why won't the clone with a mutated gene appear on an autoradiography film?
179) What are animals called that have had their DNA manipulated to possess and express an extra
(foreign) gene?
180) What percentage of all existing transgenic animals are mice?
181) Why are transgenic animals being produced?
182) Name the diseases for which transgenic models exist.
183) What are transgenic animals used for in scientific studies?
184) How can transgenic animals help in understanding the role of certain factors in the body?
185) Name an example of a complex factor involved in growth that can be studied using transgenic
animals
186) How does introducing genes from other species into transgenic animal’s aid research?
187) Why are biological products often expensive to make?
188) What is a transgenic animal?
189) Name an example of a useful biological product produced by transgenic animals
190) What was significant about Rosie, the first transgenic cow?
191) Which vaccine is currently being tested on transgenic mice for safety?
192) What is the main concern about the manipulation of living organisms by humans?
193) What role does GEAC play in India?
194) Define biopiracy.
195) How many documented varieties of Basmati rice are grown in India?
196) What did the patent rights allow the American company to do?
197) What measures are some nations taking against unauthorised exploitation of their bio-resources and
traditional knowledge?
198) Explain the role of Ti plasmids in biotechnology.
199) Assertion : RNAi is a method of cellular defence occurring in all eukaryotic organisms.
Reason : RNAi silences a specific mRNA as it binds to it to form dsRNA.
200) Assertion : GEAC has been set up to keep a check on GM research.
Reason : Genetic modification of organisms can have unpredictable results.
201) Explain the process of synthesis of insulin by Eli Lilly Company. Name the technique used by the
company.
202) Assertion : Insulin used for diabetics was earlier extracted from pancreas of live animals.
Reason : It caused allergy to some patients.
203) A person is born with a hereditary disease with a weakened immune system due to deficiency of an
enzyme. Suggest a technique for complete cure for this disease, identify the deficient enzyme and
explain the technique used for cure.
204) Plasmid is a boon to biotechnology. Justify this statement quoting the production of human insulin as
an example.
205) Give the scientific name of the soil bacterium which produces crystal (Cry) proteins. How are these
proteins useful in agriculture?
206) What do the differently written terms ‘Cry’ and ‘cry’ represent respectively?
207) Why are yeasts used extensively for functional expression of eukaryotic genes?
208) How is PCR used to detect gene mutation in case of suspected cancer patient?
209) Gene therapy is an attempt to correct a genetic defect by providing a normal gene into the
individual. By this the normal function can be restored. Alternate method would be to provide the
gene product (protein/enzyme) known as enzyme replacement therapy, which would also restore the
function. Which in your opinion is a better option? Give reason for your answer.
210) Differentiate between gene therapy and gene cloning.
211) A corn farmer has perennial problem of corn-borer infestation in his crop. Being environmentally
conscious he does not want to spray insecticides. Suggest solution based on your knowledge of
biotechnology. Write the steps to be carried out to achieve it.
212) Assertion : Agrobacterium tumefaciens is popular in genetic engineering because it spontaneously
transfers tumour inducing gene to broad leaf dicot plants.
Reason : A gene incorporated in the bacterial chromosomal genome gets autonomically transferred to
be crop with which the bacterium is associated.
213) Assertion : Genetic engineering is mainly involved in production of transgenic animals that produce
proteins.
Reason : Transgenic plants can be obtained by combination of tissue culture and genetic engineering.
Biotechnology and its Applications 435
214) Assertion : The first gene therapy was given for ADA deficiency.
Reason : The normal gene for ADA was delivered to patient’s cells using retroviral vector.
215) Assertion : Orgnisations like Genetic Engineering Approval Committee (GAEC) monitor GM
researches.
Reason : Some ethical standards are required to evaluate the morality of all human activities.
216) Assertion : Human insulin is produced in E. coli.
Reason : In mammals, insulin is synthesised as a pro-hormone which contains an extra stretch of
protein.
217) Assertion : The RNAi can be introduced in an organism only by inserting the gene encoding
complementary RNA.
Reason : The complement of the mRNA sense strand usually contains the sequence of codons for
producing functional protein.
218) Assertion : ‘Cry’ proteins are named so because they are crystal proteins.
Reason : ‘Cry’ proteins solubilise in alkaline pH of the insect’s gut and activate Bt toxin.
219) Assertion : Patents are granted by government to an inventor.
Reason : Patent prevents other from commercial use of an invention.
220) Assertion : Stem cells are undifferentiated biological cells found in multicellular organisms.
Reason : They are obtained from only umbilical cord blood just after birth.
221) Case study -
(i) How are two short polypeptide chains of insulin linked
together?
(ii) State the role of C-peptide in human insulin.
 (iii) Mention the chemical change that proinsulin undergoes, to be
able to act as mature insulin.
222) Assertion : Indian Patents Bill takes cases of biopiracy in consideration.
Reason : Biopiracy is the use of bioresources by multinational companies
without proper authorisation from concerned persons and countries.
223) Human growth hormone can be cloned and expressed experimentally
with the steps shown above.
(i) What is the host called that produces a foreign gene product?
What is this product called?
(ii) Write the name of the enzymes that are used for isolation of
DNA from bacterial and fungal cells respectively for
Recombinant DNA technology.
(iii) How can bacterial DNA be released from the bacterial cell for
biotechnology experiments?
224) What is antisense technology?
(a) When a piece of RNA that is complementary in sequence is used to stop
expression of a specific gene.
(b) RNA polymerase producing DNA.
(c) A cell displaying a foreign antigen used for synthesis of
antigens.
(d) Production of somaclonal variants in tissue cultures.
225) (a) Mark the positive and negative terminals.
(b) What is the charge carried by DNA molecule and how
does it help in its separation?
(c) How the separated DNA fragments are finally isolated?
226) (i) Name the organism in which the vector shown is inserted to get the
copies of the desired gene.
(ii) Mention the area labelled in the vector responsible for controlling the
copy number of the inserted gene.
(iii) Name and explain the role of a selectable marker in the vector
shown.
227) Assertion : Plasmids are single stranded extrachromosomal DNA.
Reason : Plasmids are found in eukaryotic cells
228) Assertion : Plasmids are extrachromosomal DNA.
Reason : Plasmids are found in bacteria and are useful in genetic
engineering.
229) Assertion : In recombinant DNA technology, human genes are often transferred into bacteria
(prokaryotes) or yeast (eukaryote).
Reason : Both bacteria and yeast multiply very fast to form huge population which express the desired
gene.
230) Assertion : Insertion of recombinant DNA within the coding sequence of beta-galactosidase results in
colourless colonies.
Reason : Presence of insert results in inactivation of enzyme beta-galactosidase known as insertional
inactivation.
231) Assertion : In recombinant DNA technology both ligase and nuclease play an important role.
Reason : Ligase cuts the DNA at specific sites and nuclease joins the DNA fragments.
232) Assertion : E.coli having pBR322 with DNA insert at BamHI site cannot grow in medium containing
tetracycline.
 Reason : Recognition site for BamHI is present in tetr region of pBR322.
233) Assertion : Downstream processing include separation and purification of product.
Reason : Before release of the product, it needs to be tested for quality control.
234) Assertion : PCR primers do not have self complementary regions.
Reason : PCR involves use of Taq polymerase as it can withstand the high temperature of the process.
235) Assertion : Restriction enzymes recognise palindromic sequences.
Reason : Palindromic sequences read the same in both directions of the strands.
236) Assertion : For isolating DNA from yeast cell, chitinase enzyme is necessary.
Reason : Cell wall of fungi are made up of chitin.

237)
238) What is known as molecular glue? Why?
239) Assertion: Transgenic food may cause toxicity or produce allergy.
Reason: Transgenic plants have high nutrient content
240) Assertion: Genetically modified microbes help in crop protection.
Reason: Transgenic bacteria control insects by producing endotoxins.
241) Assertion: Agrobacterium tumefaciens is called natural genetic engineer.
Reason: Agrobacterium tumefaciens infects all broad-leaved agricultural crops but does not infect
cereal crops.
242) Assertion: ADA deficiency cannot be cured permanently by gene therapy.
Reason: The genetically engineered lymphocytes can survive only in culture conditions.
243) Assertion: Biopatents are awarded for biological entities and all products derived from them.
Reason: Patent on use of turmeric in wound healing was cancelled in 2008.
244) Assertion: Gene therapy is a collection of methods that allows correction of a gene defect that has
been diagnosed in a child or embryo.
Reason: In gene therapy genes are inserted into a persons cells and tissues to treat a disease
245) Assertion : Transgenic mice have been engineered to express human antibodies.
Reason : Large segment of human DNA encoding human immunoglobulin have been transferred to
mice.
246) Assertion: Insulin produced by recombinant DNA technology is more effective than the insulin
produced by conventional method.
Reason: Insulin produced by recombinant DNA technology is more effectively absorbed in blood than
the insulin produced by conventional method.