Phagemid vectors — What is the most accurate description of their structure and functional advantage in cloning workflows?

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:

  • Phagemids are primarily plasmids in growth behavior.
  • An f1/M13 origin permits packaging of generated ssDNA into filamentous phage particles when a helper phage is present.
  • They are distinct from lambda, cosmid, and BAC systems.


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:

Identify key hallmark: f1/M13 origin on a plasmid backbone.Relate to function: helper phage rescue produces ssDNA for downstream applications.Choose the option that captures both structure and function.


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:

  • B: Describes lambda-based systems, not phagemids.
  • C: Incorrect biology; phages do not replicate as plasmids in eukaryotes.
  • D: Cosmids lack the f1 origin and are not phagemids.
  • E: Phagemids replicate autonomously in bacteria via plasmid ori; phage packaging is optional.


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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