Cloning in Biotechnology Quiz

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Questions and Answers

What is the primary purpose of cloning in biotechnology?

The primary purpose of cloning in biotechnology is to create genetically identical copies of organisms, cells, or DNA fragments.

Describe gene manipulation and name one method used in this process.

Gene manipulation refers to the alteration of an organism's genetic material, and one method used is gene splicing.

What role do restriction endonucleases play in the process of gene manipulation?

Restriction endonucleases are enzymes that cut DNA at specific nucleotide sequences, enabling DNA isolation.

How does DNA ligase contribute to DNA cloning?

<p>DNA ligase connects DNA fragments by joining cohesive ends, facilitating the insertion of foreign DNA.</p>
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What is a vector in molecular biology, and why is it important?

<p>A vector is a molecule of DNA that carries genetic information into cells, essential for the replication and expression of inserted DNA.</p>
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Explain the term 'transfection' and in which scenario it is used.

<p>Transfection refers to the transfer of DNA into eukaryotic cells, often used in genetic studies and therapeutic applications.</p>
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What is the significance of maintaining the ability to replicate DNA in a vector?

<p>Maintaining the ability to replicate DNA in a vector ensures that the introduced genetic material can be copied and expressed within the host.</p>
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What types of vectors are most commonly used in molecular cloning?

<p>The most commonly used vectors in molecular cloning are plasmids, viruses, and artificial chromosomes.</p>
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What is the structure of plasmids and their primary function in molecular biology?

<p>Plasmids have a double-stranded circular DNA structure, and their primary function is to carry inserted genes along with necessary enzymes and control molecules.</p>
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How is a replication origin significant for plasmids used as vectors?

<p>A replication origin is the specific DNA sequence where replication is initiated, allowing plasmids to replicate within the host.</p>
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What roles do enhancers and promoters play in gene transcription?

<p>Enhancers are regions that increase transcription likelihood by binding activators, while promoters are DNA regions that lead to the initiation of transcription.</p>
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What is a reporter gene, and why is it used in molecular biology?

<p>A reporter gene is a gene attached to a regulatory sequence of another gene, used to monitor the activity of that gene in various organisms.</p>
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Describe the role of antibiotic resistance genes in plasmids.

<p>Antibiotic resistance genes in plasmids ensure that only bacteria containing the plasmid survive antibiotic treatment, allowing researchers to select for plasmid-containing cells.</p>
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What are viral vectors, and how do they differ from plasmids?

<p>Viral vectors are artificially constructed entities containing modified DNA or RNA that can integrate into the host genome, while plasmids are circular DNA molecules that replicate independently.</p>
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What is the significance of the transcription terminator in gene expression?

<p>The transcription terminator marks the end of a gene or operon, indicating where RNA synthesis stops during transcription.</p>
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How does translation initiation occur in bacteria?

<p>Translation initiation in bacteria involves the interaction of the mRNA with the ribosomal small subunit to start protein synthesis.</p>
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Flashcards

Cloning

The process of creating identical copies of an organism, cells, or DNA fragments.

Gene Manipulation

The process of manipulating genetic material using techniques like gene splicing, recombinant DNA, or PCR.

Vector

A specific sequence of DNA that acts as a carrier of genetic information into cells for replication and expression.

Restriction Endonucleases

Enzymes that break down DNA at specific sequences, allowing scientists to isolate desired DNA fragments.

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DNA Ligase

An enzyme that joins DNA fragments together, allowing for the insertion of foreign DNA into a vector.

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Transformation (bacteria) or Transfection (eukaryotes)

The process of introducing foreign DNA into cells.

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Promoter

A molecule that binds to a specific DNA sequence, initiating the transcription of a gene.

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Agarose Gel Electrophoresis

The process of analyzing DNA fragments by size using an electric current through a gel matrix.

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Plasmids

Double-stranded circular DNA molecules that can be transferred into cells. They often carry inserted genes and necessary enzymes, allowing them to replicate and produce proteins.

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Origin of replication

A specific sequence of DNA where replication begins.

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Reporter gene

A gene used to monitor the activity of another gene in bacteria, cell culture, or living organisms.

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Enhancer

A short region of DNA that can be bound by proteins to increase the likelihood of gene transcription.

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Transcription terminator

A section of DNA sequence that signals the end of a gene during transcription.

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Transcription initiation site

The first nucleotide of a DNA sequence that starts the synthesis of an RNA transcript.

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Viral vectors

Virally derived vehicles used to deliver genetic material into cells. They are often modified to lack infectivity but retain the ability to enter and multiply within cells.

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Study Notes

Cloning in Biotechnology

  • Cloning in biotechnology refers to creating clones of organisms, cells, or DNA fragments.
  • Gene manipulation, also called genetic engineering, is a general term for methods to manipulate genetic material.

Gene Manipulation Process

  • Isolate DNA: Restriction endonucleases (type II) cut DNA at specific locations.
  • Insert into vector: DNA is joined to a vector for transport into a host cell.
  • Transfer to host: Transfection or transformation inserts the vector into host cells.

DNA Joining

  • DNA ligase joins DNA fragments with cohesive ends.
  • Agarose gel electrophoresis separates DNA fragments by size.

Vector Importance

  • Vectors carry foreign DNA into host cells for replication and expression.
  • Vectors must accept foreign DNA and continue the cell cycle.
  • Common vectors are bacteria (especially E. coli) or viruses.
  • Plasmids are common vectors with circular DNA.

Vector Types

  • Cloning vectors introduce foreign DNA into host cells and maintain it within the host organism.
    • Plasmids, cosmids, phages, BACs, YACs, or MACs are cloning vectors.
  • Expression vectors introduce DNA (gene of interest) and aid in analysing gene products (RNA or proteins).
    • Plasmids are expression vectors.
  • Both cloning and expression vectors contain: origins of replication, unique restriction sites, a reporter gene and an antibiotic resistance gene.

Insertion Methods

  • Transformation: For bacterial cells.
  • Transfection: For eukaryotic cells.
  • Transduction: For viral vectors.

Viral Vectors

  • Viral vectors contain modified DNA or RNA.
  • Viral vectors are used to transport foreign DNA to cells.
  • Common viral vectors include retroviruses.

Additional Concepts

  • Origin of replication: A specific DNA segment initiating replication.

  • Reporter gene: A gene attached to a regulatory sequence for observation.

  • Plasmids: Small, circular DNA molecules replicated independently.

  • Promoters: DNA regions initiating transcription.

  • Enhancers: DNA sequences increasing transcription likelihood.

  • Transcription terminators: DNA sequences marking the end of a gene.

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