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Question #47709
The Question
During recombinant DNA construction, a researcher uses a restriction endonuclease under suboptimal buffer conditions, resulting in incomplete cleavage. Upon agarose gel electrophoresis and Ethidium Bromide visualization, a complex banding pattern with partial digests is observed. If a DNA fragment from this gel is eluted and utilized for ligation and transformation, what is the most likely systemic consequence during the subsequent expression and downstream processing phases?
Options
- A.
The host will express truncated or chimeric proteins due to the integration of unintended sequences, compromising targeted downstream purification.
- B.
DNA ligase will fail to form phosphodiester bonds because incomplete cleavage inherently generates fragments lacking 5' phosphate groups.
- C.
The intercalated Ethidium Bromide will permanently bind to the partial digests, specifically blocking the origin of replication (ori) in the host.
- D.
Downstream processing will yield an inactive protein because the partial digestion alters the post-translational modification enzymes of the host cell.
Check Your Answer
Correct Answer
Step-by-Step Solution
Explanation
How the Examiner Thinks
Regulatory_Failure
Understand how NOT, EXCEPT, and logical constraints are used to manipulate exam results.
Concepts Behind This Question
Plasmid vs Chromosomal DNA|Restriction Enzyme Cleavage Physics|Heat Shock Thermal Gradient
Why This Question Is Tricky
Common Mistakes
Conceptual Pivot: Students often assume that any visible band on a gel is a viable cloning insert.
This question tests the ability to link an upstream molecular error (incomplete restriction digestion) to its ultimate phenotypic fallout in downstream processing.
The trap lies in distinguishing between a mechanical failure of ligation (Option B) and a failure of expression fidelity (Option A) caused by the inclusion of uncut sequences.
Quick Facts About This Question
Question Information
| Subject | Biology |
| Chapter | Biotechnology : Principles and Processes - Class 12 |
| Concept | Restriction Enzyme Cleavage Physics |
| Difficulty | 8 |
| Cognitive Load | 5 |
| Target Solve Time | 120 sec |
| Access | Free |