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Questions and Answers
What is the most critical challenge a molecular biologist faces when aiming to mass-produce a valuable protein using gene cloning techniques?
What is the most critical challenge a molecular biologist faces when aiming to mass-produce a valuable protein using gene cloning techniques?
- Identifying and isolating the specific gene that codes for the desired protein from a longer DNA molecule. (correct)
- Synthesizing the protein directly without involving genetic material.
- Creating a completely new gene sequence that enhances the protein's functionality.
- Ensuring that the bacterial plasmid is resistant to all known antibiotics.
In recombinant DNA technology, what is the primary role of bacterial plasmids?
In recombinant DNA technology, what is the primary role of bacterial plasmids?
- To degrade any foreign DNA that enters the bacteria.
- To synthesize proteins directly from the bacterial chromosome.
- To serve as vectors for carrying foreign genes into bacteria for replication and protein production. (correct)
- To provide a protective barrier for the DNA against restriction enzymes.
Which of the following steps is crucial for ensuring that a recombinant plasmid is successfully incorporated into a bacterial cell during gene cloning?
Which of the following steps is crucial for ensuring that a recombinant plasmid is successfully incorporated into a bacterial cell during gene cloning?
- Introducing a virus to inject the plasmid DNA into the bacterial cell.
- Exposing the bacteria to intense radiation to facilitate DNA uptake.
- Treating the bacteria with chemicals or electrical pulses to increase their permeability to DNA. (correct)
- Subjecting the plasmid to high temperatures to denature the DNA.
What is the primary reason for using DNA ligase in the creation of recombinant DNA?
What is the primary reason for using DNA ligase in the creation of recombinant DNA?
When cloning a gene into a bacterial plasmid, why is it important to use the same restriction enzyme to cut both the plasmid and the source DNA?
When cloning a gene into a bacterial plasmid, why is it important to use the same restriction enzyme to cut both the plasmid and the source DNA?
How does gene cloning contribute to the field of biotechnology beyond simply producing large quantities of a specific protein?
How does gene cloning contribute to the field of biotechnology beyond simply producing large quantities of a specific protein?
In the context of gene cloning, what is the most direct method to confirm that a bacterial colony contains the recombinant plasmid with the desired gene?
In the context of gene cloning, what is the most direct method to confirm that a bacterial colony contains the recombinant plasmid with the desired gene?
How do bacteria protect their own DNA from being cleaved by the restriction enzymes they produce?
How do bacteria protect their own DNA from being cleaved by the restriction enzymes they produce?
What is the major advantage of using 'sticky ends' in recombinant DNA technology?
What is the major advantage of using 'sticky ends' in recombinant DNA technology?
What is the most critical factor that determines the specificity of a restriction enzyme?
What is the most critical factor that determines the specificity of a restriction enzyme?
In the context of using nucleic acid probes, what is the primary reason for labeling the probe with a radioactive isotope or fluorescent tag?
In the context of using nucleic acid probes, what is the primary reason for labeling the probe with a radioactive isotope or fluorescent tag?
What is the most significant advantage of using DNA microarrays in genetic research?
What is the most significant advantage of using DNA microarrays in genetic research?
Why is it necessary to break open bacterial cells and separate the DNA strands before applying a nucleic acid probe in colony screening?
Why is it necessary to break open bacterial cells and separate the DNA strands before applying a nucleic acid probe in colony screening?
Which of the following best describes how a nucleic acid probe identifies bacterial colonies carrying a particular gene of interest?
Which of the following best describes how a nucleic acid probe identifies bacterial colonies carrying a particular gene of interest?
What is the main purpose of selectively breeding animals, an ancient form of biotechnology?
What is the main purpose of selectively breeding animals, an ancient form of biotechnology?
How does recombinant DNA technology differ most significantly from traditional methods of genetic manipulation, such as selective breeding?
How does recombinant DNA technology differ most significantly from traditional methods of genetic manipulation, such as selective breeding?
In the process of creating recombinant DNA, what is the role of the enzyme DNA ligase immediately after the "sticky ends" of DNA fragments have paired up?
In the process of creating recombinant DNA, what is the role of the enzyme DNA ligase immediately after the "sticky ends" of DNA fragments have paired up?
When researchers use nucleic acid probes to screen bacterial colonies, what is the most critical reason for ensuring the probe is single-stranded?
When researchers use nucleic acid probes to screen bacterial colonies, what is the most critical reason for ensuring the probe is single-stranded?
What primary advantage does the use of bacterial plasmids offer over viral vectors in gene cloning?
What primary advantage does the use of bacterial plasmids offer over viral vectors in gene cloning?
In the context of restriction enzymes, what distinguishes a 'sticky end' from a 'blunt end' and why is this difference significant for recombinant DNA technology?
In the context of restriction enzymes, what distinguishes a 'sticky end' from a 'blunt end' and why is this difference significant for recombinant DNA technology?
How does the use of nucleic acid probes in DNA microarrays fundamentally advance our understanding of genomics and proteomics?
How does the use of nucleic acid probes in DNA microarrays fundamentally advance our understanding of genomics and proteomics?
When creating recombinant DNA, why is it crucial to treat both vectors and source DNA with the same restriction enzyme?
When creating recombinant DNA, why is it crucial to treat both vectors and source DNA with the same restriction enzyme?
In colony hybridization, bacterial cells are lysed and their DNA denatured before adding a nucleic acid probe. What is the primary reason for these steps?
In colony hybridization, bacterial cells are lysed and their DNA denatured before adding a nucleic acid probe. What is the primary reason for these steps?
How do bacteria naturally prevent their DNA from being digested by their own restriction enzymes?
How do bacteria naturally prevent their DNA from being digested by their own restriction enzymes?
The use of 'sticky ends' produced by restriction enzymes is a cornerstone of recombinant DNA technology. What specific advantage do these overhanging sequences provide during the creation of recombinant molecules?
The use of 'sticky ends' produced by restriction enzymes is a cornerstone of recombinant DNA technology. What specific advantage do these overhanging sequences provide during the creation of recombinant molecules?
In DNA microarray technology, why is it crucial to use single-stranded DNA (ssDNA) rather than double-stranded DNA (dsDNA) for the probes?
In DNA microarray technology, why is it crucial to use single-stranded DNA (ssDNA) rather than double-stranded DNA (dsDNA) for the probes?
If a researcher is using a nucleic acid probe to identify a specific gene within a sample, what step should they take to increase the specificity of probe binding and minimize false positives?
If a researcher is using a nucleic acid probe to identify a specific gene within a sample, what step should they take to increase the specificity of probe binding and minimize false positives?
What is a primary advantage of using plasmids over genomic DNA when creating recombinant bacteria for protein production?
What is a primary advantage of using plasmids over genomic DNA when creating recombinant bacteria for protein production?
Flashcards
Biotechnology
Biotechnology
The manipulation of living organisms or their components to make useful products.
Recombinant DNA
Recombinant DNA
A DNA molecule that has been manipulated in the laboratory to carry nucleotide sequences derived from two sources, often different species.
Plasmid
Plasmid
A small ring of independently replicating DNA separate from the main chromosome(s). Found in prokaryotes and yeasts.
Gene cloning
Gene cloning
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Vector
Vector
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Restriction enzyme
Restriction enzyme
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Restriction site
Restriction site
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Sticky ends
Sticky ends
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DNA ligase
DNA ligase
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Nucleic acid probe
Nucleic acid probe
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Study Notes
- Biotechnology involves manipulating organisms or their components to create useful products.
- DNA technology refers to modern lab techniques used for studying and manipulating genetic material.
- Recombinant DNA methods combine DNA from different sources to create a single DNA molecule in a lab.
- Recombinant DNA technology is used in genetic engineering to manipulate genes for practical applications.
- Bacterial plasmids are small, circular DNA molecules that replicate independently of the bacterial chromosome.
- Plasmids are key tools for DNA cloning, which produces many identical copies of a target DNA segment.
- Gene cloning enables the mass production of useful products.
Gene Cloning Process
- Isolate bacterial plasmid (vector) and foreign DNA containing the desired gene.
- Treat both plasmid and source DNA with a restriction enzyme that cuts DNA at specific sites.
- Mix the cut DNA from the plasmid and source, allowing complementary single-stranded ends to base-pair.
- Use DNA ligase to join the DNA molecules, creating a recombinant DNA molecule.
- Introduce the recombinant plasmid into bacteria through transformation.
- The recombinant bacterium reproduces, forming a clone of genetically identical cells, each carrying a copy of the desired gene.
Purposes of Gene Cloning
- Producing copies of the gene.
- Harvesting the protein product.
- Cloned genes can be used in further genetic engineering projects.
- Recombinant bacteria can be used to produce medical proteins like insulin in large quantities.
Enzymes in DNA Manipulation
- Restriction enzymes act as cutting tools, recognizing specific short DNA sequences called restriction sites.
- Restriction enzymes cut both DNA strands at precise points, creating restriction fragments.
- DNA ligase joins DNA fragments by forming covalent bonds between adjacent nucleotides.
Restriction Enzymes
- Restriction enzymes are bacterial enzymes that cut up foreign DNA as a defense mechanism.
- Bacteria protect their own DNA by adding methyl groups.
- Restriction fragments are the pieces of cut DNA.
Sticky Ends
- Sticky ends are single-stranded extensions from double-stranded DNA fragments produced by the same restriction enzyme.
- Sticky ends are complementary and can stick together by base pairing.
- Complementary ends on DNA fragments stick together by base pairing, allowing them to be joined.
Recombinant DNA Formation
- Recombinant DNA is formed by joining restriction fragments from different sources.
- Sticky ends play a key role in this process, as hydrogen bonds form base pairs that hold the strands together.
- DNA ligase then makes the union between fragments permanent.
Nucleic Acid Probes
- Researchers can use a known part of a nucleotide sequence to find a specific DNA segment.
- A nucleic acid probe is a synthesized short single strand of DNA with a complementary sequence to the target DNA.
- Nucleic acid probes are labeled with a radioactive isotope or fluorescent tag to make them detectable.
- The labeled probe binds to the complementary sequence in the target DNA.
Function of Radioactive DNA Probes
- When a probe is added to a collection of DNA strands, it tags the correct molecules by hydrogen-bonding to the complementary sequence in the gene of interest.
- This allows researchers to screen DNA molecules simultaneously for a desired gene.
Applications of Nucleic Acid Probes
- Used in DNA microarrays to test the expression of multiple genes simultaneously.
- Nucleic acid probes allow researchers to identify bacterial colonies carrying a gene of interest.
- After identification, the cells of the tagged colony can be grown further.
- The gene of interest, or its protein product, can be collected in large amounts.
Terms to Know
- Biotechnology: The manipulation of living organisms or their components to make useful products.
- Recombinant DNA: A DNA molecule manipulated in the laboratory to carry nucleotide sequences from two sources.
- Vector: A piece of DNA used to move genes from one cell to another.
- Restriction enzymes: Bacterial enzymes that cut up foreign DNA at specific DNA sequences.
- Restriction site: Specific DNA sequence where a restriction enzyme cuts.
- Plasmid: A small ring of independently replicating DNA separate from the main chromosome.
- Gene cloning: The production of multiple copies of a gene.
- Nucleic acid probe: A labeled single-stranded nucleic acid molecule used to find a specific gene or nucleotide sequence within a mass of DNA.
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