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Advances in biotechnology are possible because of key techniques such as
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Restriction enzyme analysis
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Blotting techniques
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DNA sequencing
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solid- phase synthesis of amino acids
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solid-phase synthesis of nucleic acids
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PCR
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Restriction enzymes split DNA into specific fragments
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- True
- False
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Restriction enzymes are
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bacterial enzymes that cleave DNA in a highly specific manner
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fungal enzymes that cleave DNA in a highly specific manner
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phage enzymes that cleave DNA in a highly specific manner
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The restriction enzymes recognize
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cleavage sites of three to eight nucleotides in length that are palindromic and then cleave each strand of the DNA
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cleavage sites of four to eight nucleotides in length that are palindromic and then cleave each strand of the DNA
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cleavage sites of four to eight nucleotides in length that are non-palindromic and then cleave each strand of the DNA
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cleavage sites of three to eight nucleotides in length that are non-palindromic and then cleave each strand of the DNA
Frage 5
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DNA fragments can be separated by gel electrophoresis
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- True
- False
Frage 6
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In gel electrophoresis, fragments can be visualized by staining with
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agarose
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polyacrylamide gels
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ethidium bromide
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bromide
Frage 7
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Separated fragments of DNA are transferred to a sheet of
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cellulose paper, a process called southern blotting, and then exposed to a radioactively labeled probe complementary to the sequence of interest
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nitrocellulose paper, a process called southern blotting, and then exposed to a radioactively labeled probe parallel to the sequence of interest
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nitrocellulose paper, a process called southern blotting, and then exposed to a radioactively labeled probe complementary to the sequence of interest
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cellulose paper, a process called southern blotting, and then exposed to a radioactively labeled probe parallel to the sequence of interest
Frage 8
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Autoradiography does not identify the fragment binding to the probe
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- True
- False
Frage 9
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The polymerase chain reaction (PCR)
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Allows the amplification of any DNA sequence provided some sequence information about the target DNA is known
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Allows the amplification of any DNA sequence without the sequence information about the target DNA
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Separates DNA fragments
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Consist of target DNA and does not require heat in order to stabilize the DNA polymerase
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PCR consist of three steps that are repeated until the desired degree of amplification is achieved. What is the first step?
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The mixture is cooled to a temperature determined by the experimenter (55 degrees Celsius), to allow the primers to anneal to the DNA
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The duplex containing the target sequence and the flanking regions (primers) is cooled to 80 degrees to generate single strands
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The duplex containing the target sequence and the flanking regions (primers) is heated to 80 degrees Celsius to generate single strands
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The mixture is heated to 72 degrees Celsius, allowing the polymerase to synthesize DNA
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PCR is a powerful technique in medical diagnosis, forensics, and studies of molecular evolution
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- True
- False
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Instances of genetic variation
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Polymorphisms, does not correlate with the emergence of a disease
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Polymorphisms, sometimes correlate with the emergence of a disease
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Polymorphisms, sometimes correlate with the disappearance of a disease
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Polymorphisms, does not correlate with the disappearance of a disease
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A vector,
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a piece of RNA readily taken up and replicate by bacteria, is cleaved by the same restriction enzyme used to generate the fragment
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a piece of DNA readily taken up and replicate by bacteria, is cleaved by the same restriction enzyme used to generate the fragment
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a piece of RNA ligase readily taken up and replicate by bacteria, is cleaved by the same restriction enzyme used to generate the fragment
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a piece of DNA ligase readily taken up and replicate by bacteria, is cleaved by the same restriction enzyme used to generate the fragment
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DNA ligase is used to separate the fragment and the vector, generating recombinant DNA
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- True
- False
Frage 15
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Common Vectors are
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DNA ligase
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bacterial plasmids
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bacteriophage lambda
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primers
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If the DNA fragment of interest does not contain appropriate restriction sites for vector insertion, a chemically synthesized linker DNA with the required restriction site is added to the fragment
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- True
- False
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Plasmids and lamda phage are choice vectors for DNA cloning in bacteria
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Specifically designed plasmids called cloning vectors allow for efficient deletion of DNA as well as replication
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Specifically designed plasmids called cloning vectors allow for efficient insertion of DNA as well as replication
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cloning vectors may not have a polylinker region
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cloning vectors may have a polylinker regions
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cloning Vectors contain promoters that facilitate transcription of the insert and sequences that allow translation
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cloning Vectors contain promoters that facilitate transcription of the deletion and sequences that allow translation
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Reporter genes, such as antibiotic-resistance genes, in the vector make identification of vectors with the inserted DNA harder
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- True
- False
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Lamda phage (viruses that infect bacteria) are
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cloning vectors
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can infect a cell and cause lysis or become stably integrated into the host DNA (lysogenic pathway) and be replicated indefinitely
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Environmental changes does not cause the switch from lysogenic to lytic pathway
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complementary DNA prepared from mRNA
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Complementary DNA can also be inserted into expression vectors
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- True
- False
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Expression vectors are
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plasmids or λ phage that have powerful promoters for transcription as well as a segment of DNA that encodes a ribosome-binding site that is expressed with the mRNA encoded by the cDNA.
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plasmids or λ phage that have powerful promoters for translation as well as a segment of DNA that encodes a ribosome-binding site that is expressed with the rRNA encoded by the cDNA.
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plasmids or λ phage that have powerful promoters for transcription as well as a segment of DNA that encodes a ribosome-binding site that is expressed with the tRNA encoded by the cDNA.
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plasmids or λ phage that have powerful promoters for translation as well as a segment of DNA that encodes a ribosome-binding site that is expressed with the tRNA encoded by the cDNA.
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Expression vectors allow for not only transcription but also translation of the cDNA library
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- True
- False
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Replica plates are made of a λ phage containing an expression vector, and the plaques are probed with an antibody for the protein of interest.
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- True
- False
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Human gene therapy holds great promise for medicine
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Gene therapy, the deletion of functional genes into cells to replace mutated genes
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Gene therapy, the insertion of functional genes into cells to replace mutated genes
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Severe combined immunodeficiency (SCID)
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Is an example of gene therapy
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is an example of a phage
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has been treated by infecting cells with a functional version of the mutated enzyme, adenosine deaminase
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has been treated by curing cells with a functional version of the mutated enzyme, adenosine deaminase
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