Biotechnology Quiz: Recombinant DNA Technology

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Questions and Answers

What is the primary purpose of a marker gene?

To identify transformed organisms (correct)

To provide resistance to pests

To enhance growth rates in plants

To produce antibiotics

Which gene provides resistance to Tetracycline?

gfp

tetr (correct)

ampr

What application does recombinant DNA technology have in agriculture?

Production of insulin

Development of new antibiotics

Manufacture of Bt Cotton (correct)

Creation of antibody probes

Which of the following is NOT a concern related to recombinant DNA technology?

Resistance to pests in crops Signup and view all the answers

What major breakthrough was achieved in 1982 regarding insulin?

Human insulin was produced in engineered bacteria Signup and view all the answers

What distinguishes a transformed cell from a competent cell?

Transformed cells have taken up foreign DNA successfully Signup and view all the answers

What characteristic does the gfp gene provide to GloFish?

Fluorescence under UV light Signup and view all the answers

Which of the following is an advantage of recombinant DNA technology?

Unlimited utilizations Signup and view all the answers

What is the initial step in the PCR process?

Denaturation Signup and view all the answers

Which component is NOT always required in a PCR reaction?

Dye Signup and view all the answers

At what temperature does the elongation phase of PCR occur?

72°C Signup and view all the answers

How many cycles are typically conducted in a PCR process?

25-40 Signup and view all the answers

What is the primary role of primers in a PCR reaction?

To serve as a starting point for DNA replication Signup and view all the answers

What occurs during the annealing step of PCR?

DNA primers bind to the template DNA Signup and view all the answers

What can cause a PCR amplification reaction to fail?

Presence of co-extracted inhibitors Signup and view all the answers

What is the temperature range typically used for primer annealing in PCR?

55°-65°C Signup and view all the answers

What is the role of DNA ligase in recombinant DNA technology?

It joins DNA fragments with cloning vectors. Signup and view all the answers

Why is the efficiency of ligation low when using blunt ends of DNA?

Blunt ends lack complementary sequences for binding. Signup and view all the answers

What is the function of terminal transferase in DNA manipulation?

It synthesizes a short sequence of complementary nucleotides. Signup and view all the answers

What is the purpose of alkaline phosphatase in recombinant DNA technology?

To prevent self-annealing of vector DNA. Signup and view all the answers

What defines a competent cell in the context of transformation?

A cell that can take up DNA from its environment. Signup and view all the answers

Which enzyme is responsible for adding a phosphate group to the terminal 5' end of DNA?

Polynucleotide kinase Signup and view all the answers

What is the main purpose of restriction endonucleases in DNA manipulation?

To cut DNA at specific sequences. Signup and view all the answers

What critical function does methylation provide to a host cell's DNA in the presence of endonucleases?

It protects the host DNA from degradation. Signup and view all the answers

What is the primary function of the buffer in gel electrophoresis?

To provide ions that carry a current and maintain pH Signup and view all the answers

Which type of gel concentration is suitable for resolving small pieces of DNA (50–500 bp)?

2% – 3% agarose gel Signup and view all the answers

How does the size of a DNA fragment affect its migration in gel electrophoresis?

Smaller fragments travel further than larger fragments Signup and view all the answers

What is the role of ethidium bromide in DNA fragment separation?

To intercalate between DNA base pairs and enhance visibility under UV light Signup and view all the answers

What is the outcome after 30 cycles of a standard PCR reaction?

Approximately 1 billion copies of DNA Signup and view all the answers

What characteristic of Taq polymerase makes it suitable for PCR?

It is thermostable and can withstand high temperatures Signup and view all the answers

What must be designed specifically for the region of interest in a PCR reaction?

DNA primers Signup and view all the answers

Why is it important that molecules to be separated in electrophoresis must have charges?

The electric current requires charge to move the molecules Signup and view all the answers

What is the first step in the DNA extraction process?

Lysis Signup and view all the answers

Which enzyme is specifically used to destroy RNA during DNA extraction?

RNAase Signup and view all the answers

What is the purpose of washing the DNA pellet with ethanol?

To purify the DNA Signup and view all the answers

Which method can be used to assess the purity of extracted DNA?

Measuring absorbance at 260 and 280 nm Signup and view all the answers

In gel electrophoresis, what primarily determines the migration of DNA molecules?

The size and charge of the molecules Signup and view all the answers

What type of DNA extraction sample can be used?

Animal or tissue samples Signup and view all the answers

What is the expected A260/A280 ratio for pure DNA?

∼1.8 Signup and view all the answers

What are the two main parts of most DNA extraction protocols?

Lysis and precipitation Signup and view all the answers

Study Notes

Recombinant DNA Technology

Involves selecting a desired gene and a suitable vector for integration into a host.
DNA ligase joins DNA fragments with cloning vectors.
Restriction endonucleases recognize specific sequences (restriction sites) to cut DNA.
Endonucleases are isolated from various microorganisms.
Terminal transferase converts blunt ends of DNA fragments into sticky ends.
DNA polymerase synthesizes complementary nucleotides.
Alkaline phosphatase removes terminal phosphate groups.
Polynucleotide kinase adds phosphate groups to terminal 5' ends.
Recombinant DNA technology has five basic steps: culturing host cells, extracting products, downstream processing of the products.
Gene splicing involves cutting DNA at a specific site.
Vectors carry foreign genes into host cells.
Transformation is the introduction of free DNA into bacteria.
Competent cells can take up DNA.
Electroporation uses an electric shock to disrupt cell walls in order to allow DNA uptake.
Transformed cells have new DNA.
Marker genes identify successfully transformed organisms.
pBR322 is a plasmid vector.
tet gene confers resistance to tetracycline.
amp gene confers resistance to ampicillin.

Applications of Recombinant DNA Technology

Recombinant proteins are used in laboratory experiments and to generate antibodies.
Bacteria can produce human insulin and growth hormone.
Bacteria can be engineered to consume oil spills.
Recombinant DNA technology has applications in agriculture.

DNA Extraction

DNA extraction can be done from nucleated cells.
The goal of DNA extraction obtains a relatively pure form for further analysis.
DNA extraction procedures have two main parts: lysing the cell and removing contaminants.
The technique involves: lysis, precipitation, wash, re-suspension
Detergents are used to disrupt the plasma membrane.
Proteases and RNAases are used to eliminate proteins and RNA.
Supernatant (containing DNA) is transferred to a clean tube and is precipitated using ethanol.
Methods include the use of detergent/enzymes, centrifugation, and precipitation.

Gel Electrophoresis

Separates molecules based on charge and size in a gel.
Charged molecules migrate in an electrical field through a gel matrix.
Gel's porosity affects migration speed.
Molecules migrate towards the opposite charge.
Used for separating DNA fragments and other biological molecules.
Gel electrophoresis uses buffers (e.g., TAE, TBE) to maintain constant pH and support current flow.
DNA is negatively charged.
Smaller pieces travel further than larger ones.

PCR (Polymerase Chain Reaction)

PCR amplifies specific DNA sequences.
PCR uses a thermostable DNA polymerase (e.g., Taq polymerase) and specific primers.
PCR steps include denaturation, annealing, and extension.
Denaturation creates single-stranded DNA.
Primers anneal to the template DNA.
Taq polymerase extends the primers along the template DNA.

Blotting Techniques

Transferring DNA, RNA, or proteins from a gel to a membrane for analysis.
Southern blotting detects DNA.
Northern blotting detects RNA.
Western blotting detects proteins.
Techniques are helpful to study mutations in genes
Advantages: good sensitivity, multiple protein analysis, and detection of mutations.

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Description

Test your knowledge on recombinant DNA technology and its applications in biotechnology. This quiz covers topics such as marker genes, gene resistance, and the significant breakthroughs in insulin production. Determine your understanding of how these technologies impact agriculture and the environment.