Insertion vectors (lambda): What is the maximum practical size of foreign DNA that can be cloned using a λ insertion vector?

Difficulty: Medium

Correct Answer: About 10 kb

Explanation:


Introduction / Context:
Lambda vectors come in two major formats for cloning: insertion vectors and replacement vectors. Insertion vectors retain most of the λ genome and accept relatively small inserts. Replacement vectors remove a stuffer fragment to accommodate much larger inserts. Knowing these capacities is crucial when selecting the right vector for a given fragment size.



Given Data / Assumptions:

  • Packaging constraints require the total recombinant DNA to fit within the λ headful window.
  • Insertion vectors accept small inserts; replacement vectors accept much larger inserts.
  • Common insertion vectors (for example, λ gt10) typically handle inserts up to roughly the 7–10 kb range.


Concept / Approach:
Because insertion vectors keep most of the λ genome intact, only a limited additional size can be added while keeping the total within packaging limits. In practice, inserts much above ~10 kb are not suitable for insertion vectors and should be moved to replacement formats.



Step-by-Step Solution:

Confirm vector class → insertion type retains bulk of λ DNA.Apply packaging window → little room left for extra DNA.Select the approximate maximum → around 10 kb insert size.


Verification / Alternative check:
Cloning guides for λ insertion vectors recommend inserts in the vicinity of several kilobases, and commonly state inefficiency beyond ~10 kb, whereas replacement vectors target ~15–20+ kb up to ~40 kb.



Why Other Options Are Wrong:

35 kb or 50 kb: typical of replacement or cosmid systems, not insertion vectors.18 kb: exceeds practical insertion-vector capacity.< 2 kb only: too restrictive; insertion vectors can clone several kb effectively.


Common Pitfalls:
Attempting large genomic fragments in insertion vectors and obtaining very low packaging or plaque yields.



Final Answer:
About 10 kb.

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