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Biology 2 - C01 - 04
🧬🧩BIO02 Module 1 — Composite MasterSet — 04
| Question | Answer |
|---|---|
| What is genetic engineering? | The deliberate modification of an organism’s DNA using biotechnology. |
| What is recombinant DNA technology? | A method of combining DNA from different sources to create new genetic combinations. |
| What is a vector? | A DNA carrier (often a plasmid or virus) used to deliver foreign genes into a host cell. |
| What is a plasmid? | A small, circular DNA molecule used as a common cloning vector. |
| What is a transgenic organism? | An organism that contains DNA from another species. |
| What do restriction enzymes do? | They cut DNA at specific recognition sequences. |
| What are sticky ends? | Single-stranded overhangs created by staggered restriction enzyme cuts. |
| What are blunt ends? | Straight DNA cuts with no overhangs. |
| What does DNA ligase do? | It seals DNA fragments by forming phosphodiester bonds. |
| What does reverse transcriptase do? | It synthesizes complementary DNA (cDNA) from an mRNA template. |
| What is Step 1 of recombinant DNA technology? | DNA isolation from donor and host organisms. |
| What is Step 2 of recombinant DNA technology? | Cutting DNA with restriction enzymes to create compatible ends. |
| What is Step 3 of recombinant DNA technology? | Ligation of foreign DNA into a vector. |
| What is Step 4 of recombinant DNA technology? | Transformation of recombinant DNA into host cells. |
| What is Step 5 of recombinant DNA technology? | Selection and screening of transformed cells. |
| What is transformation? | The uptake of foreign DNA by a host cell. |
| What is heat-shock transformation? | A method using temperature changes to make bacterial membranes permeable. |
| What is electroporation? | Using electric pulses to open pores in cell membranes for DNA entry. |
| What is a selectable marker? | A gene used to identify transformed cells, often antibiotic resistance. |
| What is blue-white screening? | A method using lacZ disruption to distinguish recombinant from non-recombinant colonies. |
| What is gene therapy? | The insertion of functional genes to treat genetic disorders. |
| What is CRISPR-Cas9? | A gene-editing tool using guide RNA and Cas9 nuclease to cut DNA precisely. |
| What is a GMO? | An organism whose genome has been altered using genetic engineering. |
| How is insulin produced using recombinant DNA? | Human insulin gene is inserted into bacteria, which express the protein. |
| What is DNA fingerprinting? | A technique using STRs to identify individuals. |
| What is biosafety level (BSL)? | A classification of lab safety protocols based on organism risk. |
| What is bioethics? | The study of ethical issues arising from biotechnology. |
| What is the Cartagena Protocol? | An international agreement regulating the movement of GMOs. |
| What is gene drive? | A genetic system that increases the likelihood of a gene being inherited. |
| What are off-target effects? | Unintended DNA modifications caused by gene-editing tools. |
| What is biotechnology? | The use of living systems or organisms and technology to develop products or processes for practical purposes. |
| What is cloning? | Creating genetically identical copies of DNA, cells, or whole organisms; can occur naturally (identical twins) or be done artificially (e.g., somatic cell nuclear transfer) for research, agriculture, or therapeutic purposes. |
| What is DNA sequencing? | Determining the exact order of nucleotides (A, T, C, G) in a DNA molecule to read genes or whole genomes, used in research, diagnostics, evolutionary studies, and forensics. |
| What is Polymerase Chain Reaction (PCR)? | A method using repeated heating/cooling cycles and DNA polymerase to amplify a specific DNA fragment. |
| What is genome editing? | Techniques (e.g., CRISPR) that make precise, targeted changes to an organism’s DNA sequence. |
| 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. |
| Purpose of Recombinant DNA Technology | The goal is to combine DNA from different sources to create new genetic combinations for research, medicine, or biotechnology. |
| Recognition Sequence (Restriction Enzymes) | A short, specific DNA sequence (often palindromic) where a restriction enzyme cuts the DNA. |
| Viral Vector | A virus modified to carry foreign DNA into host cells for gene delivery or expression. |
| Origin of Replication (Ori) | A DNA sequence in a vector that allows it to replicate independently inside a host cell. |
| Selectable Marker | A gene in a vector (e.g., antibiotic resistance) that allows identification of cells that successfully took up the vector. |
| Multiple Cloning Site (MCS) | A short DNA region containing many restriction enzyme sites, allowing easy insertion of foreign DNA. |
| Reporter Gene | A gene that produces a visible or measurable product (e.g., GFP, LacZ) to indicate successful gene expression or cloning. |
| PCR: Denaturation | The step where DNA strands separate when heated to around 94–98°C. |
| PCR: Annealing | The step where primers bind to target DNA at 50–65°C. |
| PCR: Extension | The step where Taq polymerase synthesizes new DNA strands at around 72°C. |
| Role of Taq Polymerase | A heat‑stable enzyme that builds new DNA strands during PCR. |
| Applications of PCR | Used for DNA amplification, diagnostics, forensics, cloning, and detecting pathogens. |
| How DNA Separates in Gel Electrophoresis | DNA fragments move through a gel toward the positive electrode; smaller fragments travel farther. |
| Role of Agarose in Electrophoresis | Agarose forms a porous gel matrix that separates DNA fragments by size. |
| Interpreting Gel Bands | Each band represents DNA fragments of a specific size; distance traveled indicates fragment length. |
| cDNA vs Genomic DNA (Difference) | cDNA contains only expressed genes (no introns), while genomic DNA includes introns and non‑coding regions. |
| When cDNA Is Used | Used when expressing eukaryotic genes in bacteria or studying gene expression. |
| Cloning Vector | A vector designed to carry and replicate foreign DNA but not necessarily express it. |
| Expression Vector | A vector that includes regulatory sequences to ensure the inserted gene is transcribed and translated into protein. |
| Role of Reverse Transcriptase | An enzyme that converts mRNA into complementary DNA (cDNA). |
| Screening vs Selection | Selection kills or inhibits non‑transformed cells; screening visually distinguishes colonies without killing them. |
| Example of Genetic Engineering: Bt Corn | Corn engineered to express Bt toxin, giving resistance to insect pests. |
| Example of Genetic Engineering: Insulin Production | Bacteria engineered with the human insulin gene to produce insulin for medical use. |
Created by:
frankie gabriel slavin