Question:
Which of the following is the activity of alkaline phosphatase?
(1). Addition of 5'-\( \text{PO}_4^{3-} \)
(2). Removal of 5'-\( \text{PO}_4^{3-} \)
(3). Removal of nucleotides from 3'-ends
(4). Removal of single-strand protrusions from the end
Which of the following is the activity of alkaline phosphatase?
(1). Addition of 5'-\( \text{PO}_4^{3-} \)
(2). Removal of 5'-\( \text{PO}_4^{3-} \)
(3). Removal of nucleotides from 3'-ends
(4). Removal of single-strand protrusions from the end
(1). Addition of 5'-\( \text{PO}_4^{3-} \)
(2). Removal of 5'-\( \text{PO}_4^{3-} \)
(3). Removal of nucleotides from 3'-ends
(4). Removal of single-strand protrusions from the end
Show Hint
Remember:
Kinase \( \rightarrow \) Adds phosphate.
Phosphatase \( \rightarrow \) Removes phosphate.
Alkaline phosphatase is used to prevent plasmid self-ligation in cloning.
Kinase \( \rightarrow \) Adds phosphate.
Phosphatase \( \rightarrow \) Removes phosphate.
Alkaline phosphatase is used to prevent plasmid self-ligation in cloning.
Updated On: Jul 31, 2026
- Addition of 5'-\( \text{PO}_4^{3-} \)
- Removal of 5'-\( \text{PO}_4^{3-} \)
- Removal of nucleotides from 3'-ends
- Removal of single-strand protrusions from the end
Show Solution
Verified By Collegedunia
The Correct Option is B
Solution and Explanation
Step 1: Understanding the Concept:
Phosphatases are hydrolase enzymes that remove phosphate groups from organic molecules.
Alkaline phosphatase operates optimally in alkaline pH environments and is a standard tool used in recombinant DNA technology.
Step 2: Detailed Explanation:
Let us examine the specific actions of the listed options:
1. Removal of 5'-Phosphate (2): Alkaline phosphatase catalyzes the removal of 5'-phosphate groups from DNA, RNA, and ribo/deoxyribonucleoside triphosphates.
In molecular cloning, treating linearized vector DNA with alkaline phosphatase (such as Calf Intestinal Phosphatase, CIP, or Shrimp Alkaline Phosphatase, SAP) removes the 5' phosphate group required by DNA ligase.
This prevents the vector from self-ligating (re-circularizing) without an insert, reducing background colonies.
2. Addition of 5'-Phosphate (1): This phosphorylation reaction is catalyzed by T4 Polynucleotide Kinase (PNK), which transfers a phosphate from ATP to a 5'-OH group.
3. Removal of nucleotides (3 and 4): Exonucleases or single-strand-specific nucleases (like Mung Bean Nuclease or S1 Nuclease) are responsible for removing nucleotides or single-strand overhangs.
Therefore, the correct enzymatic activity of alkaline phosphatase is the removal of 5'-phosphate groups.
Step 3: Final Answer:
Alkaline phosphatase removes the 5'-phosphate group, which corresponds to option (B).
Phosphatases are hydrolase enzymes that remove phosphate groups from organic molecules.
Alkaline phosphatase operates optimally in alkaline pH environments and is a standard tool used in recombinant DNA technology.
Step 2: Detailed Explanation:
Let us examine the specific actions of the listed options:
1. Removal of 5'-Phosphate (2): Alkaline phosphatase catalyzes the removal of 5'-phosphate groups from DNA, RNA, and ribo/deoxyribonucleoside triphosphates.
In molecular cloning, treating linearized vector DNA with alkaline phosphatase (such as Calf Intestinal Phosphatase, CIP, or Shrimp Alkaline Phosphatase, SAP) removes the 5' phosphate group required by DNA ligase.
This prevents the vector from self-ligating (re-circularizing) without an insert, reducing background colonies.
2. Addition of 5'-Phosphate (1): This phosphorylation reaction is catalyzed by T4 Polynucleotide Kinase (PNK), which transfers a phosphate from ATP to a 5'-OH group.
3. Removal of nucleotides (3 and 4): Exonucleases or single-strand-specific nucleases (like Mung Bean Nuclease or S1 Nuclease) are responsible for removing nucleotides or single-strand overhangs.
Therefore, the correct enzymatic activity of alkaline phosphatase is the removal of 5'-phosphate groups.
Step 3: Final Answer:
Alkaline phosphatase removes the 5'-phosphate group, which corresponds to option (B).
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