25. The restriction enzymes Kpnl and Acc65l 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' GGTACC3 3 CCATGGS 3CCATGGS 5' GGTACC3 Kpnl Acc651 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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Chapter1: The Human Body: An Orientation
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### Transcription and Explanation:

**Question 25:**
The restriction enzymes KpnI and Acc65I recognize and cleave the same 6-base pair sequence. You have a plasmid and a linear DNA strand that both contain a KpnI and Acc65I 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?

**Diagram Explanation:**
The diagram depicts the cleavage sites for KpnI and Acc65I.

- **KpnI and Acc65I Recognition Sites:**
  - Both enzymes recognize the sequence 5' GGTACC 3' on one strand and 3' CCATGG 5' on the complementary strand.
  - The cleavage occurs as indicated by the arrows, separating the DNA into sticky ends.

**Answer Options:**

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:### Transcription and Explanation: **Question 25:** The restriction enzymes KpnI and Acc65I recognize and cleave the same 6-base pair sequence. You have a plasmid and a linear DNA strand that both contain a KpnI and Acc65I 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? **Diagram Explanation:** The diagram depicts the cleavage sites for KpnI and Acc65I. - **KpnI and Acc65I Recognition Sites:** - Both enzymes recognize the sequence 5' GGTACC 3' on one strand and 3' CCATGG 5' on the complementary strand. - The cleavage occurs as indicated by the arrows, separating the DNA into sticky ends. **Answer Options:** 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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