Difficulty: Easy
Correct Answer: Ligase
Explanation:
Introduction:
DNA manipulation often requires sealing phosphodiester backbone breaks. The canonical enzyme used in cloning and in vivo repair is being asked for here.
Given Data / Assumptions:
Concept / Approach:
DNA ligases form a ligase–AMP intermediate and transfer AMP to the 5'-phosphate, activating the end for nucleophilic attack by the 3'-OH, thus sealing the nick. In cloning, T4 DNA ligase is standard; cells have their own ligases for replication and repair.
Step-by-Step Solution:
Verification / Alternative check:
Standard cloning protocols rely on T4 DNA ligase to join cohesive or blunt ends, confirming the role and nomenclature.
Why Other Options Are Wrong:
Common Pitfalls:
Assuming any enzyme that touches DNA can join fragments; only ligases perform the final sealing step in cloning.
Final Answer:
Ligase.
Discussion & Comments