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24. Sickle-cell anemia arises from a mutation in the gene for the B chain of human hemoglobin. The change from GAG to GTG in the mutant that is located within the cleavage site for the restriction enzyme Mstll, which recognizes the target sequence CCTGAGG. These findings form the basis of a diagnostic test for the sickle-cell gene. Which answer best explains how this test should work? A) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the normal gene but not the mutant gene. B) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the mutant gene but not the normal gene. C) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the normal gene. D) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the mutant gene. E) Restriction enzymes are non-specific relative to the binding requirements for such a test.

24. Sickle-cell anemia arises from a mutation in the gene for the B chain of human hemoglobin. The change from GAG to GTG in the mutant that is located within the cleavage site for the restriction enzyme Mstll, which recognizes the target sequence CCTGAGG. These findings form the basis of a diagnostic test for the sickle-cell gene. Which answer best explains how this test should work? A) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the normal gene but not the mutant gene. B) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the mutant gene but not the normal gene. C) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the normal gene. D) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the mutant gene. E) Restriction enzymes are non-specific relative to the binding requirements for such a test.

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
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24. Sickle-cell anemia arises from a mutation in the gene for the B chain of human hemoglobin. The change from GAG to GTG in the mutant that is located within the cleavage site for the restriction enzyme Mstll, which recognizes the target sequence CCTGAGG. These findings form the basis of a diagnostic test for the sickle-cell gene. Which answer best explains how this test should work? A) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the normal gene but not the mutant gene. B) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the mutant gene but not the normal gene. C) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the normal gene. D) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the mutant gene. E) Restriction enzymes are non-specific relative to the binding requirements for such a test. 25. The restriction enzymes Kpnl and Acc651 recognize and cleave the same 6-bp sequence. You have a plasmid and a linear DNA strand that both contain a Kpnl and Acc651 sequence in the same orientation as shown below. You digest both DNA pieces with both enzymes and then attempt to ligate the sticky ends, followed by treatment with DNA ligase. What will happen? 5' GGTACC 3' 5' GGTACC3' 3' CCATGGS 3 CCATGGS Kpnl Acс651 A) You will produce sticky ends but the two types of ends will not ligate. Instead, you may produce a small amount of religated plasmid where the digested plasmid sequence re-inserts. B) You will produce a recombinant plasmid in which the linear DNA strand is ligated in between the two sites, suitable for cloning. C) You will produce blunt ends that will not ligate because the two restriction enzymes will both operate on both of the sites. D) All the DNA will be completely digested as if you had applied a general DNAse enzyme.
Transcribed Image Text:24. Sickle-cell anemia arises from a mutation in the gene for the B chain of human hemoglobin. The change from GAG to GTG in the mutant that is located within the cleavage site for the restriction enzyme Mstll, which recognizes the target sequence CCTGAGG. These findings form the basis of a diagnostic test for the sickle-cell gene. Which answer best explains how this test should work? A) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the normal gene but not the mutant gene. B) In combination with DNA polymerase, application of Mstll will produce long DNA strands with the mutant gene but not the normal gene. C) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the normal gene. D) Treatment with Mstll produces DNA fragments from the normal gene but cannot cleave the mutant gene. E) Restriction enzymes are non-specific relative to the binding requirements for such a test. 25. The restriction enzymes Kpnl and Acc651 recognize and cleave the same 6-bp sequence. You have a plasmid and a linear DNA strand that both contain a Kpnl and Acc651 sequence in the same orientation as shown below. You digest both DNA pieces with both enzymes and then attempt to ligate the sticky ends, followed by treatment with DNA ligase. What will happen? 5' GGTACC 3' 5' GGTACC3' 3' CCATGGS 3 CCATGGS Kpnl Acс651 A) You will produce sticky ends but the two types of ends will not ligate. Instead, you may produce a small amount of religated plasmid where the digested plasmid sequence re-inserts. B) You will produce a recombinant plasmid in which the linear DNA strand is ligated in between the two sites, suitable for cloning. C) You will produce blunt ends that will not ligate because the two restriction enzymes will both operate on both of the sites. D) All the DNA will be completely digested as if you had applied a general DNAse enzyme.
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