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Biology 2 - C01 - 02

🧬📗BIO02 Module 1 Genetic Engineering and Recombinant DNA Technology 002

TermDefinition
Define genetic engineering. The deliberate modification of an organism’s DNA using biotechnology.
What is a vector in recombinant DNA technology? A DNA carrier (often a plasmid or virus) used to deliver foreign genes into a host cell.
What are sticky ends? Single-stranded overhangs created by staggered restriction enzyme cuts.
Define transformation. The uptake of foreign DNA by a host cell.
What is reverse transcriptase used for? Converting mRNA into complementary DNA (cDNA).
Arrange the steps: Transformation, DNA isolation, Screening, Ligation, Restriction digestion. DNA isolation → Restriction digestion → Ligation → Transformation → Screening.
Why must restriction enzymes create compatible ends? To allow the foreign DNA and vector to base-pair correctly for ligation.
What is the role of DNA ligase? It seals DNA fragments by forming phosphodiester bonds.
Why is heat-shock used in bacterial transformation? The temperature change increases membrane permeability, allowing DNA entry.
Compare electroporation and heat-shock. Electroporation uses electric pulses; heat-shock uses temperature shifts.
How is recombinant DNA used to produce human insulin? The human insulin gene is inserted into bacterial plasmids; bacteria express the protein.
Why is blue-white screening useful? White colonies indicate successful insertion of foreign DNA; blue colonies do not.
Give one medical and one agricultural application of genetic engineering. Medical: gene therapy or insulin production. Agricultural: pest-resistant or herbicide-tolerant crops.
Why use cDNA instead of genomic DNA in cloning? cDNA lacks introns, making it expressible in bacteria.
How can CRISPR-Cas9 correct a mutation? gRNA guides Cas9 to the mutation site; Cas9 cuts; repair template fixes the sequence.
Restriction sites don’t match—give two solutions. Use different restriction enzymes OR add linkers/adaptors to create compatible ends.
Why are antibiotic resistance genes used as selectable markers? Only transformed cells survive on antibiotic plates, making identification easy.
How do off-target CRISPR effects pose risks? They may unintentionally cut other genes, causing mutations or harmful effects.
What do white colonies indicate in lacZ screening? The foreign DNA disrupted lacZ, meaning successful recombinant plasmids.
Why is recombinant DNA technology both powerful and ethically sensitive? It enables major advances but raises concerns about safety, equity, and unintended consequences.
 

 



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