Difficulty: Medium
Correct Answer: Hybrid plasmids carrying a filamentous phage origin (f1/M13) to allow single-stranded DNA rescue
Explanation:
Introduction / Context:
Phagemids merge convenient plasmid features (ori, antibiotic resistance, MCS) with a filamentous phage origin of replication (f1/M13). This enables recovery of single-stranded DNA after helper phage infection—critical for Sanger sequencing, mutagenesis, and phage display scaffolds.
Given Data / Assumptions:
Concept / Approach:
A phagemid replicates as a double-stranded plasmid under normal growth. Upon superinfection with a helper phage, the f1 ori directs production of circular ssDNA that is secreted in filamentous particles. This duality combines ease of manipulation with the sequencing advantages of ssDNA templates.
Step-by-Step Solution:
Verification / Alternative check:
Vector manuals for pBluescript, pGEM, and similar phagemids describe helper phage rescue protocols to obtain ssDNA for Sanger sequencing or mutagenesis.
Why Other Options Are Wrong:
Common Pitfalls:
Confusing cosmid packaging (dsDNA) with filamentous phage ssDNA rescue; they serve very different purposes.
Final Answer:
Hybrid plasmids carrying a filamentous phage origin (f1/M13) to allow single-stranded DNA rescue
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