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Biotechnology
Genetic Engineering

Genetic engineering, fermentation, cell biology

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Topics in Biotechnology
During the development of a genetically modified organism (GMO), scientists use selectable marker genes. If an antibiotic resistance gene is used as a marker, which scenario would indicate successful transformation?
A Transformed cells survive on media containing the antibiotic; non-transformed cells die
B Non-transformed cells survive on media containing the antibiotic; transformed cells die
C Both transformed and non-transformed cells survive equally on antibiotic media
D Neither transformed nor non-transformed cells survive on antibiotic media
Correct Answer:  A. Transformed cells survive on media containing the antibiotic; non-transformed cells die
EXPLANATION

Selectable marker genes confer resistance to specific antibiotics or toxins. When the transformation vector carries an antibiotic resistance gene, only cells that successfully incorporated the vector will express the resistance gene and survive on antibiotic-containing media. Non-transformed cells lack this gene and are eliminated.

Test
In CRISPR-Cas9 gene editing technology, what is the specific role of the single guide RNA (sgRNA)?
A It synthesizes new DNA strands after Cas9 cuts the DNA
B It directs the Cas9 nuclease to the target DNA sequence by complementary base pairing
C It repairs double-strand breaks created by Cas9
D It prevents off-target cutting by blocking non-specific sites
Correct Answer:  B. It directs the Cas9 nuclease to the target DNA sequence by complementary base pairing
EXPLANATION

The sgRNA contains a CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA) combined into a single molecule. It guides the Cas9 nuclease to the target DNA sequence through Watson-Crick base pairing, ensuring specificity of the cut.

Test
A biotechnology company in India has developed a transgenic crop that produces its own Bt toxin for pest resistance. Which of the following best explains the mechanism by which this toxin controls insect pests?
A It inhibits protein synthesis in insect gut cells by breaking down peptide bonds
B It creates pores in the insect midgut epithelial membrane, leading to cell lysis and death
C It interferes with the insect nervous system by blocking neurotransmitter receptors
D It reduces the insect's ability to metabolize plant sugars
Correct Answer:  B. It creates pores in the insect midgut epithelial membrane, leading to cell lysis and death
EXPLANATION

Bt toxin (Cry proteins) forms pores in the alkaline midgut of target insects, causing cell lysis and disruption of the intestinal epithelium. This is specific to lepidopteran, coleopteran, and dipteran insects with appropriate pH and protease conditions.

Test
Which of the following enzymes is primarily used to cut DNA at specific recognition sequences during genetic engineering?
A Restriction endonuclease
B DNA polymerase
C Helicase
D Ligase
Correct Answer:  A. Restriction endonuclease
EXPLANATION

Restriction endonucleases (restriction enzymes) recognize and cut DNA at specific palindromic sequences, forming sticky or blunt ends. DNA polymerase synthesizes DNA, helicase unwinds DNA, and ligase joins DNA strands.

Test
In the production of recombinant human insulin using genetically engineered bacteria, which organism is primarily used?
A Bacillus subtilis
B Escherichia coli (E. coli)
C Salmonella typhimurium
D Streptococcus pneumoniae
Correct Answer:  B. Escherichia coli (E. coli)
EXPLANATION

E. coli is the primary organism used for recombinant insulin production due to its well-characterized genetics, rapid growth, and ability to produce high levels of recombinant proteins.

Test
Which of the following represents a limitation of homologous recombination-based gene targeting?
A Very high efficiency in mammalian cells without selection
B Low frequency of homologous recombination in mammalian somatic cells (~1 in 10^6-10^7)
C Inability to create specific mutations
D High off-target effects
Correct Answer:  B. Low frequency of homologous recombination in mammalian somatic cells (~1 in 10^6-10^7)
EXPLANATION

Homologous recombination has very low frequency in mammalian cells compared to yeast, requiring enrichment strategies and selection markers. This limitation has been overcome by CRISPR technology.

Test
In next-generation sequencing (NGS), what is the primary advantage of whole-genome sequencing over targeted gene sequencing?
A Lower cost per base
B Detection of unknown mutations and structural variations across the entire genome
C Faster processing time
D Higher accuracy for specific genes
Correct Answer:  B. Detection of unknown mutations and structural variations across the entire genome
EXPLANATION

Whole-genome sequencing provides unbiased coverage of the entire genome, allowing discovery of unexpected mutations, copy number variations, and structural rearrangements not detected by targeted approaches.

Test
What is the primary mechanism of action of TALENs (Transcription Activator-Like Effector Nucleases) in gene editing?
A Formation of secondary structures to block transcription
B Modular DNA-binding domain directs FokI nuclease to specific target sequences
C Competitive inhibition of RNA polymerase
D Methylation of target DNA
Correct Answer:  B. Modular DNA-binding domain directs FokI nuclease to specific target sequences
EXPLANATION

TALENs consist of a modular DNA-binding domain (which recognizes specific sequences) fused to the FokI nuclease domain, allowing precise DNA cleavage at target sites.

Test
Which Indian government body is primarily responsible for approving the release of genetically modified crops?
A Ministry of Environment, Forest and Climate Change (MoEFCC)
B Genetic Engineering Appraisal Committee (GEAC)
C Indian Council of Medical Research (ICMR)
D Department of Biotechnology (DBT) only
Correct Answer:  B. Genetic Engineering Appraisal Committee (GEAC)
EXPLANATION

The GEAC, operating under the MoEFCC, is responsible for appraising large-scale field trials and commercial release of genetically modified organisms in India.

Test
In the context of RNA interference (RNAi), what is the role of the RISC complex?
A To amplify target mRNA
B To recognize and cleave complementary mRNA with guide RNA
C To transcribe DNA
D To synthesize new RNA
Correct Answer:  B. To recognize and cleave complementary mRNA with guide RNA
EXPLANATION

The RNA-Induced Silencing Complex (RISC) uses small RNA molecules (siRNA or miRNA) as guides to recognize and degrade complementary mRNA sequences, leading to gene silencing.

Test
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