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.

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.

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.

A gene that produces a visible or measurable product (e.g., GFP, LacZ) to indicate successful gene expression or cloning.

The step where DNA strands separate when heated to around 94–98°C.

The step where primers bind to target DNA at 50–65°C.

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.

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.

Used when expressing eukaryotic genes in bacteria or studying gene expression.

A vector designed to carry and replicate foreign DNA but not necessarily express it.

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.

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Biology 2 - C01 - 03

🧬📗1️⃣ Module 1 Genetic Engineering and Recombinant DNA Technology 003

Question	Answer
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

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