Expression from λ gt11: How is an inserted DNA fragment typically expressed when cloned into the lambda gt11 vector?

Difficulty: Easy

Correct Answer: As a β-galactosidase fusion protein from the lacZ gene

Explanation:


Introduction / Context:
λ gt11 is a classic expression cloning vector used to produce fusion proteins for immunoscreening. The vector is designed so that inserted DNA is fused in-frame with the lacZ gene, producing a β-galactosidase fusion polypeptide. This enables plaque immunoscreening with antibodies against the fusion partner.



Given Data / Assumptions:

  • λ gt11 contains the lac promoter and the 5’ portion of lacZ.
  • Cloning in-frame into the multiple cloning site yields a LacZ–insert fusion.
  • Fusion proteins can be detected with anti-β-galactosidase or insert-specific antibodies.


Concept / Approach:
The vector architecture forces expression as a β-galactosidase fusion if the insert is properly oriented and in-frame. This facilitates expression screening and identification of antigenic clones directly on plates.



Step-by-Step Solution:

Recognize vector design → lacZ-based fusion system.Determine expression product → β-galactosidase fusion protein with the cloned insert.Eliminate alternatives that do not match the vector mechanism (free or secreted proteins).


Verification / Alternative check:
Protocols for λ gt11 immunoscreening emphasize detecting LacZ–fusion antigens in plaques using specific antisera.



Why Other Options Are Wrong:

Free cytosolic or secreted protein: Not the default outcome of λ gt11 design.All of the above/membrane-anchored: Not supported by the vector's expression strategy.


Common Pitfalls:
Frame-shifted inserts that abolish fusion; cloning in the wrong orientation; using inappropriate host strains lacking necessary lac expression components.



Final Answer:
As a β-galactosidase fusion protein from the lacZ gene.

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