Q1. Match the following cloning vector components with their roles:
Column I
Column II
A. ori
(I) Identifying non-transformants
B. Selectable marker
(II) Initiation of replication
C. Ligase
(III) Joining DNA ends
Correct Answer: (a)
Standard definitions of vector components (Page 9).
Q2. Which of the following is correct regarding 'Restriction Enzymes'? (i) Belong to a larger class called nucleases. (ii) Hind II was the first one isolated. (iii) They cut only one strand of the DNA double helix.
Correct Answer: (a)
Page 6: They cut each of the two strands of the double helix.
Q3. Identify true statements for 'Selectable Markers': (i) Help in identifying transformants. (ii) Help in eliminating non-transformants. (iii) Genes for antibiotic resistance are commonly used.
Correct Answer: (a)
Role of markers from Section 9.2.2.
Q4. Which of the following have the ability to replicate within bacterial cells independent of chromosomal DNA control?
Correct Answer: (c)
Page 8: Plasmids and bacteriophages have the ability to replicate within bacterial cells independent of the control of chromosomal DNA.
Q5. Assertion: Palindromes in DNA read the same on both strands when orientation is kept same.
Reason: EcoRI recognizes the sequence GAATTC.
Correct Answer: (a)
Page 6: Both are true facts about palindromes and EcoRI, but the example doesn't define the concept.
Q6. Which enzyme is used to remove RNA during the isolation of DNA?
Correct Answer: (b)
Page 11: The RNA can be removed by treatment with ribonuclease.
Q7. Match the following vector types:
A. BAC
Bacterial Artificial Chromosome
B. YAC
Yeast Artificial Chromosome
Correct Answer: (a)
Vectors used in HGP (Page 25).
Q8. Which of the following is correct? (i) Herbert Boyer studied restriction enzymes in E. coli. (ii) Stanley Cohen studied plasmids. (iii) They met in Hawaii in 1969.
Correct Answer: (a)
Biographical history from Page 2.
Q9. The specific DNA sequence in a chromosome which is responsible for initiating replication is called:
Correct Answer: (c)
Page 4: In a chromosome there is a specific DNA sequence called the origin of replication, which is responsible for initiating replication.
Q10. In 1972, the first recombinant DNA was constructed by linking an antibiotic resistance gene with a native plasmid of:
Correct Answer: (b)
Page 4: Stanley Cohen and Herbert Boyer accomplished this in 1972 using the native plasmid of Salmonella typhimurium.
Q11. Which class of enzymes do restriction enzymes belong to?
Correct Answer: (b)
Page 6: Restriction enzymes belong to a larger class of enzymes called nucleases.
Q12. The process of extracting separated bands of DNA from the agarose gel is called:
Correct Answer: (b)
Page 8: The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. This step is known as elution.
Q13. Regarding the 'First Recombinant DNA' (1972): (i) Constructed by Stanley Cohen and Herbert Boyer. (ii) Involved Salmonella typhimurium plasmid. (iii) Used DNA ligase to link genes.
Correct Answer: (a)
Historical details from Section 9.1.
Q14. The enzyme used to break open bacterial cells to release DNA is:
Correct Answer: (c)
Page 11: This can be achieved by treating the bacterial cells... with enzymes such as lysozyme.
Q15. Assertion: DNA ligase is referred to as 'molecular scissors'.
Reason: It helps in joining the cut ends of DNA molecules.
Correct Answer: (a)
Page 5: Restriction enzymes are scissors; Page 4: Ligases are the glue (joining ends).
Q16. Match the PCR steps with their functions:
Step
Purpose
A. Denaturation
(I) DNA polymerase adds nucleotides
B. Annealing
(II) Primers bind to template
C. Extension
(III) Separating DNA strands using heat
Correct Answer: (a)
PCR mechanism (Page 12).
Q17. Protease is used during DNA isolation to remove:
Correct Answer: (c)
Page 11: whereas proteins can be removed by treatment with protease.
Q18. Assertion: Traditional hybridization often leads to inclusion of undesirable genes.
Reason: Genetic engineering allows the isolation and introduction of only desirable genes.
Correct Answer: (b)
Page 4: Both are true facts, but the limitation of one is simply the advantage of the other; the reason doesn't explain why hybridization includes undesirable ones.
Q19. The 'rop' gene in vector pBR322 codes for:
Correct Answer: (b)
Page 9: rop codes for the proteins involved in the replication of the plasmid.
Q20. Assertion: Bioprocess engineering involves maintaining a sterile ambience.
Reason: Sterile conditions allow the growth of multiple types of microbes in a bioreactor.
Correct Answer: (a)
Page 4: Sterile ambience is to enable growth of only the desired microbe.
Q21. Purified DNA precipitates out after the addition of:
Correct Answer: (b)
Page 11: Purified DNA ultimately precipitates out after the addition of chilled ethanol.
Q22. How many restriction enzymes have been isolated from over 230 strains of bacteria?
Correct Answer: (c)
Page 5: Today we know more than 900 restriction enzymes that have been isolated from over 230 strains of bacteria.
Q23. Waiters on the 'sticky ends' of DNA form ______ bonds with their complementary counterparts.
Correct Answer: (c)
Page 7: These are named so because they form hydrogen bonds with their complementary cut counterparts.
Q24. Which of the following is true for 'Transformation'? (i) It is a procedure to introduce alien DNA into a host. (ii) Bacterial cells must be made competent using calcium. (iii) It is achieved by incubating cells with DNA on ice, followed by 42°C heat shock.
Correct Answer: (a)
Complete process described in Section 9.2.3.
Q25. Restriction enzymes are also known as:
Correct Answer: (b)
Page 5: Restriction enzymes are referred to as 'molecular scissors'.
Q26. Match the scientist with their discovery:
Column I
Column II
A. Herbert Boyer
(I) Plasmids
B. Stanley Cohen
(II) Restriction enzymes / Sticky ends
C. Frederick Sanger
(III) Amino acid sequencing
Correct Answer: (a)
Biographical info on Page 2 and 25.
Q27. Which of the following is considered a selectable marker for E. coli?
Correct Answer: (b)
Page 9: the genes encoding resistance to antibiotics such as ampicillin... are considered useful selectable markers for E. coli.
Q28. The first restriction endonuclease to be isolated and characterized was:
Correct Answer: (c)
Page 5: The first restriction endonuclease—Hind II... was isolated and characterised five years later.
Q29. Identify the correct steps in 'Recombinant DNA Technology': (i) Isolation of DNA. (ii) Ligation of DNA fragment into a vector. (iii) Culturing host cells at large scale. (iv) Downstream processing.
Correct Answer: (a)
Full sequence given in Section 9.3.
Q30. Assertion: Recombinant DNA can be forced into a bacterium by heat shock at 42°C.
Reason: Heat shock makes the cell wall impermeable to DNA.
Correct Answer: (c)
Page 11: Heat shock is a method to enable the bacteria to take up DNA, not make it impermeable.
Q31. Identify correct facts for 'Ethidium Bromide': (i) Used to stain DNA in gel. (ii) Shows bright orange bands. (iii) Requires UV light for visualization.
Correct Answer: (a)
Visualisation protocol from Page 8.
Q32. The process including separation and purification of the product is collectively referred to as:
Correct Answer: (b)
Page 15: The processes include separation and purification, which are collectively referred to as downstream processing.
Q33. Match the restriction enzyme with its source organism:
Enzyme
Source
A. EcoRI
(I) Haemophilus influenzae
B. Hind II
(II) Escherichia coli
Correct Answer: (a)
Nomenclature rules (Page 5).
Q34. Identify correct features of the 'Sparged Stirred-tank Bioreactor': (i) Sterile air bubbles are sparged. (ii) Dramatic increase in oxygen transfer area. (iii) No agitator system.
Correct Answer: (a)
Page 14: Sparging increases area; most stirred-tanks have agitators.
Q35. Assertion: DNA cannot pass through cell membranes.
Reason: DNA is a hydrophilic molecule.
Correct Answer: (a)
Page 10: Since DNA is a hydrophilic molecule, it cannot pass through cell membranes.
Q36. The overhanging single-stranded portions left after restriction enzyme action are called:
Correct Answer: (b)
Page 7: There are overhanging stretches called sticky ends on each strand.
Q37. To make bacterial cells 'competent' to take up DNA, they are treated with a specific concentration of a divalent cation such as:
Correct Answer: (b)
Page 10: This is done by treating them with a specific concentration of a divalent cation, such as calcium.
Q38. The primary role of the stirrer in a stirred-tank bioreactor is to:
Correct Answer: (b)
Page 14: The stirrer facilitates even mixing and oxygen availability throughout the bioreactor.
Q39. Which of the following statements about 'Bioreactors' is true? (i) They provide optimal growth conditions (temp, pH, salts). (ii) The most common type is the stirred-tank. (iii) They process 1 to 10 litres of culture.
Correct Answer: (a)
Page 14: Bioreactors process large volumes (100-1000 litres).
Q40. Assertion: Downstream processing involves separation and purification.
Reason: It is done before the product is ready for marketing.
Correct Answer: (a)
Page 15: The steps ensure the product is finished and pure for market entry.
Q41. Which method of gene transfer is suitable for plants using high-velocity micro-particles of gold or tungsten?
Correct Answer: (b)
Page 11: In another method, suitable for plants, cells are bombarded with high velocity micro-particles... in a method known as biolistics or gene gun.
Q42. Hind II always cuts DNA molecules at a particular point by recognizing a specific sequence of:
Correct Answer: (b)
Page 5: Hind II always cut DNA molecules at a particular point by recognising a specific sequence of six base pairs.
Q43. Which of the following is correct? (i) DNA replication results into polyploidy if cell division fails. (ii) Biotechnology products include antibiotics, vaccines, enzymes. (iii) DNA is a hydrophilic molecule.
Correct Answer: (a)
General facts from the text (Page 13, 3, 10).
Q44. Assertion: In gel electrophoresis, smaller DNA fragments move farther than larger ones.
Reason: The agarose gel provides a sieving effect based on size.
Correct Answer: (a)
Page 8: The DNA fragments separate according to their size through sieving effect... smaller the fragment size, the farther it moves.
Q45. The DNA in a prokaryotic cell is found in a region termed as:
Correct Answer: (b)
Page 5: DNA (being negatively charged) is held with some proteins... in a region termed as ‘nucleoid’.
Q46. The European Federation of Biotechnology (EFB) definition of biotechnology encompasses:
Correct Answer: (c)
Page 3: The EFB definition encompasses both traditional view and modern molecular biotechnology.
Q47. Select correct statements for 'DNA Polymerases': (i) Use a DNA template to catalyse polymerisation. (ii) Highly efficient and fast. (iii) Rate in E. coli is 2000 bp per second.
Correct Answer: (a)
Enzyme properties from Section 5.4.2 (referenced in Unit IX).
Q48. Assertion: The 'ori' sequence is responsible for controlling the copy number of linked DNA.
Reason: If one wants many copies, the target DNA should be cloned in a vector with an origin supporting high copy number.
Correct Answer: (a)
Page 9: The reason provides the functional application of the assertion.
Q49. Assertion: Ethidium bromide stained gels are exposed to UV light for visualization.
Reason: Purified DNA fragments can be seen clearly in visible light without staining.
Correct Answer: (a)
Page 8: You cannot see pure DNA fragments in the visible light and without staining.
Q50. Match the Following:
A. $\beta$-galactosidase
Enzyme for lactose hydrolysis
B. Taq polymerase
Thermostable DNA polymerase
Correct Answer: (a)
Enzymes from Section 9.3.
Q51. Correct features of 'DNA Polymorphism': (i) Basis of genetic mapping. (ii) Variation at genetic level arises due to mutations. (iii) Frequency must be greater than 0.01 in human population.
Correct Answer: (a)
Definitions from Section 5.10 (referenced in Unit IX).
Q52. Match the following bioreactor types:
Column I
Column II
A. Simple stirred-tank
(I) Sterile air bubbles sparged
B. Sparged stirred-tank
(II) Agitator system
Correct Answer: (a)
Figure 9.7 labels (Page 14).
Q53. Match the Following:
Term
Description
A. Palindrome
(I) Overhanging DNA stretches
B. Sticky ends
(II) Reads same forward/backward
C. Elution
(III) Extraction of DNA from gel
Correct Answer: (a)
Standard molecular biology terms (Page 6-8).
Q54. Identify the correct statements for 'Gel Electrophoresis': (i) Agarose is a natural polymer from sea weeds. (ii) DNA fragments separate according to their size. (iii) Smaller fragments move slower through the gel.
Correct Answer: (a)
Page 8: The smaller the fragment size, the farther (faster) it moves.
Q55. DNA fragments are negatively charged and can be separated by forcing them to move towards the:
Correct Answer: (b)
Page 8: Since DNA fragments are negatively charged molecules they can be separated by forcing them to move towards the anode under an electric field.
Q56. Which pathogen is used as a vector to deliver 'T-DNA' to transform normal plant cells into a tumor?
Correct Answer: (b)
Page 10: Agrobacterium tumifaciens... is able to deliver a piece of DNA known as ‘T-DNA’ to transform normal plant cells into a tumor.
Q57. The three steps of each cycle in PCR are:
Correct Answer: (b)
Page 12, Figure 9.6: Each cycle has three steps: (i) Denaturation; (ii) Primer annealing; and (iii) Extension of primers.
Q58. Regarding 'HGP' methodologies: (i) ESTs focus on identifying genes expressed as RNA. (ii) Sequence Annotation involves sequencing all coding and non-coding parts. (iii) BAC and YAC were used as vectors.
Correct Answer: (a)
HGP methods described on Page 25.
Q59. In which method is recombinant DNA directly injected into the nucleus of an animal cell?
Correct Answer: (c)
Page 11: In a method known as micro-injection, recombinant DNA is directly injected into the nucleus of an animal cell.
Q60. Which of the following phylum/group is being described by the given statements? (i) These are autonomously replicating circular extra-chromosomal DNA. (ii) They are used to multiply alien DNA numbers. (iii) Stanley Cohen and Herbert Boyer used them in 1972.
Correct Answer: (b)
Description of plasmids from Section 9.1 and 9.2.2.
Q61. Assertion: Chilled ethanol is used at the final step of DNA isolation.
Reason: DNA precipitates out as fine threads upon addition of ethanol.
Correct Answer: (a)
Page 11: Ethanol addition causes the precipitation, making the DNA visible.
Q62. Match the Following:
Term
Unit
A. Bioreactor volume
100-1000 litres
B. PCR amplification
1 billion copies
Correct Answer: (a)
Data from Page 14 and 13.
Q63. The most commonly used matrix in gel electrophoresis is agarose, which is a natural polymer extracted from:
Correct Answer: (b)
Page 8: Nowadays the most commonly used matrix is agarose which is a natural polymer extracted from sea weeds.
Q64. Match the following gene transfer methods:
Column I
Column II
A. Micro-injection
(I) Plant cells (gold/tungsten)
B. Biolistics
(II) Animal cell nucleus
C. Disarmed pathogen
(III) Agrobacterium / Retrovirus
Correct Answer: (a)
Methods of introducing alien DNA (Page 11).
Q65. In the nomenclature of EcoRI, the letter 'R' is derived from the name of the:
Correct Answer: (c)
Page 5: In EcoRI, the letter ‘R’ is derived from the name of strain (RY 13).
Q66. Match the following for DNA charge and movement:
Term
Description
A. DNA Charge
Negative
B. Movement in Electrophoresis
Towards Anode (positive)
Correct Answer: (a)
Page 8 descriptions.
Q67. Assertion: Insertional inactivation of $\beta$-galactosidase is used for selection of recombinants.
Reason: Recombinants produce blue coloured colonies in the presence of chromogenic substrate.
Correct Answer: (a)
Page 10: recombinants... do not produce any colour (Non-recombinants produce blue colour).
Q69. Which statements regarding 'DNA Isolation' are true? (i) Proteins removed by protease. (ii) RNA removed by ribonuclease. (iii) Purified DNA is seen as fine threads in chilled ethanol.
Correct Answer: (a)
Standard isolation protocol (Page 11).
Q70. Assertion: The DNA polymerase used in PCR is isolated from Thermus aquaticus.
Reason: It is a thermostable enzyme.
Correct Answer: (a)
Page 13: Thermostability is the specific quality required for PCR cycles.
Q71. The thermostable DNA polymerase used in PCR is isolated from the bacterium:
Correct Answer: (c)
Page 13: repeated amplification is achieved by the use of a thermostable DNA polymerase (isolated from a bacterium, Thermus aquaticus).
Q72. Correct statements about 'Sticky Ends' include: (i) Facilitate the action of DNA ligase. (ii) Form hydrogen bonds with complementary counterparts. (iii) Produced by exonucleases.
Correct Answer: (a)
Page 7: They are produced by endonucleases like EcoRI.
Q73. DNA fragments separated in a gel can be visualized only after staining with:
Correct Answer: (c)
Page 8: The separated DNA fragments can be visualised only after staining the DNA with a compound known as ethidium bromide.
Q74. Match the following for the lac operon genes:
Column I (Gene)
Column II (Product)
A. z gene
(I) Permease
B. y gene
(II) Beta-galactosidase
C. a gene
(III) Transacetylase
Correct Answer: (a)
Page 22: z (beta-gal), y (permease), a (transacetylase).
Q75. Assertion: A plasmid is autonomously replicating circular extra-chromosomal DNA.
Reason: Plasmids are always found in all eukaryotic cells.
Correct Answer: (a)
Page 4: Plasmids are found in bacteria (like Salmonella). They are not standard in all eukaryotes.
Q76. The genetic material of all organisms without exception is:
Correct Answer: (d)
Page 11: Recall that nucleic acid is the genetic material of all organisms without exception.
Q77. Identify the correct phylum/group being described: (i) It is a sequence from where replication starts. (ii) Controls the copy number of linked DNA. (iii) Essential for any vector.
Correct Answer: (c)
Description of 'ori' from Section 9.2.2.
Q78. Which of the following is correct regarding 'Insertional Inactivation'? (i) Used to differentiate recombinants from non-recombinants. (ii) Based on the ability to produce color with chromogenic substrate. (iii) Recombinants produce blue color.
Correct Answer: (a)
Page 10: Recombinants... do not produce any colour.
Q79. Match the following for transformation markers:
A. Insertional inactivation
Selection of recombinants
B. Chromogenic substrate
Color-based selection
Correct Answer: (a)
Alternative markers (Page 10).
Q80. Assertion: Stanley Cohen and Herbert Boyer founded the discipline of biotechnology.
Reason: They were the first to recombine segments of DNA and insert them into bacterial cells.
Correct Answer: (a)
Page 2: Their breakthrough was the basis upon which the discipline was founded.
Q81. Regarding 'Ti Plasmid': (i) Obtained from Agrobacterium tumifaciens. (ii) Modified into a cloning vector. (iii) Still pathogenic to plants.
Correct Answer: (a)
Page 10: Modified Ti plasmid is no more pathogenic to plants.
Q82. Assertion: A vector must have very few recognition sites.
Reason: Multiple recognition sites within a vector generate several fragments, complicating gene cloning.
Correct Answer: (a)
Page 9: Minimal sites are required to avoid fragmentation.
Q83. Assertion: Bioreactors have a foam control system.
Reason: Continuous agitation and aeration in a bioreactor can lead to foam formation.
Correct Answer: (a)
Page 14: Bioreactors are designed with systems to handle process-byproducts like foam.
Q84. Regarding 'pBR322', which statements are correct? (i) It is an E. coli cloning vector. (ii) It has antibiotic resistance genes ampR and tetR. (iii) 'rop' codes for the proteins involved in replication.
Correct Answer: (a)
Full description provided on Page 9 (Figure 9.4).
Q85. Match the following for HGP:
Organism
Base pairs
A. E. coli
(I) 3.3 x 10^9 bp
B. Human (Haploid)
(II) 4.6 x 10^6 bp
Correct Answer: (a)
Data from Page 80 (Unit VII context in Unit IX).
Q86. The presence of a chromogenic substrate gives ______ coloured colonies if the plasmid does not have an insert.
Correct Answer: (b)
Page 10: The presence of a chromogenic substrate gives blue coloured colonies if the plasmid in the bacteria does not have an insert.
Q87. Which are correct for 'Biolistics'? (i) Also called gene gun. (ii) Uses gold or tungsten micro-particles. (iii) Recombinant DNA is directly injected into the nucleus.
Correct Answer: (a)
Page 11: Direct injection is micro-injection, not biolistics.
Q88. Which are correct for 'Palindromes' in DNA? (i) Read same on both strands when orientation is kept same. (ii) EcoRI sequence is 5'--GAATTC--3'. (iii) Read same as word-palindromes (e.g. MALAYALAM).
Correct Answer: (a)
Page 6: Word palindromes read same in both directions; DNA palindromes read same on two different strands in same orientation.
Q89. If any protein encoding gene is expressed in a heterologous host, it is called a:
Correct Answer: (b)
Page 13: If any protein encoding gene is expressed in a heterologous host, it is called a recombinant protein.
Q90. The average size of a human cell DNA is 2.2 metres, while the dimension of a typical nucleus is approximately:
Correct Answer: (b)
Page 5: A length that is far greater than the dimension of a typical nucleus (approximately 10^–6 m).
Q91. Regarding 'PCR', identify the correct statements: (i) Used for amplification of gene of interest. (ii) Primers are small chemically synthesised oligonucleotides. (iii) Taq polymerase is sensitive to high temperatures.
Correct Answer: (a)
Page 13: Taq polymerase is thermostable (remains active during high temp).
Q92. Assertion: Thermus aquaticus is used in PCR.
Reason: It provides a DNA polymerase that remains active during high-temperature denaturation.
Correct Answer: (a)
Page 13: repeated amplification is achieved by the use of a thermostable DNA polymerase (Taq polymerase) from T. aquaticus.
Q93. To produce large quantities of biotechnological products, vessels called ______ are used.
Correct Answer: (b)
Page 14: the development of bioreactors, where large volumes (100-1000 litres) of culture can be processed, was required.
Q94. Enzymes that remove nucleotides from the ends of the DNA are called:
Correct Answer: (b)
Page 6: Exonucleases remove nucleotides from the ends of the DNA.
Q95. Match the enzymes in Column I with their target substrates in Column II:
Q96. The specific sequence of base pairs that reads the same on the two strands when orientation is kept the same is called:
Correct Answer: (b)
Page 6: 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.
Q97. Select correct statements for 'Downstream Processing': (i) Includes separation and purification. (ii) Product is formulated with preservatives. (iii) It is done before biosynthetic stage.
Correct Answer: (a)
Page 14: It is done after the completion of the biosynthetic stage.
Q98. Which technique enabled the birth of modern biotechnology by allowing the alteration of genetic material?
Correct Answer: (b)
Page 3: Genetic engineering is one of the two core techniques that enabled the birth of modern biotechnology.
Q99. Assertion: Agrobacterium tumifaciens is known as 'nature's genetic engineer'.
Reason: It naturally delivers 'T-DNA' to transform normal plant cells into tumor cells.
Correct Answer: (a)
Page 10: The natural ability to transfer DNA makes it a tool for scientists.
Q100. Assertion: Recombination is the generation of non-parental gene combinations.
Reason: Linkage describes the physical association of genes on a chromosome.
Correct Answer: (a)
Page 17 (Unit VII context in Unit IX): Both are valid definitions but independent.
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