Molecular cloning — What core features define an expression vector used to express a cloned gene in a host cell (consider replication, selection, and transcriptional control)?

Difficulty: Easy

Correct Answer: All of the above

Explanation:


Introduction:
An expression vector is a specialized cloning vector designed not merely to carry DNA but to express it as RNA and protein inside a chosen host (for example, Escherichia coli, yeast, or mammalian cells). This question tests whether you can identify the essential architectural features that make expression possible and selectable.


Given Data / Assumptions:

  • The host must replicate the plasmid to inherit it.
  • Researchers must select cells that actually contain the plasmid.
  • Transcription and often translation require appropriate regulatory DNA sequences.


Concept / Approach:
Evaluate each proposed feature as part of a minimal, practical expression system. A workable vector must be replicated (origin), selected (antibiotic or auxotrophic marker), and expressed (promoter plus elements like ribosome-binding site or Kozak sequence). The multiple-cloning site alone is insufficient.


Step-by-Step Solution:

1) Origin of replication: enables autonomous plasmid replication in the host (e.g., ColE1-derived ori for E. coli).2) Selectable marker: allows survival on a selective medium (e.g., ampicillin, kanamycin), ensuring maintenance of the vector in culture.3) Expression control: a promoter suited to the host (e.g., T7, lac, CMV), plus additional elements such as operator, RBS/Kozak, terminator/poly(A) for eukaryotes, and often tags for purification.4) Therefore, a, b, and c are all correct; the comprehensive answer is “All of the above.”


Verification / Alternative check:
Commercial expression vectors (pET, pGEX, pcDNA) universally include ori + selectable marker + promoter/terminator modules, illustrating the standard design principles.


Why Other Options Are Wrong:

e) A cloning site without ori/marker/regulatory elements cannot support stable expression or selection.


Common Pitfalls:
Confusing general cloning vectors with expression vectors; forgetting that promoters are host-specific (a bacterial promoter will not drive expression in mammalian cells without the correct eukaryotic elements).


Final Answer:
All of the above.

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