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In which of the given circumstances it is challenging to differentiate between recombinant and non-recombinant cells based on the insertion of foreign DNA at specific recognition sites in the pBR322 vector?
Option: 1
When the foreign DNA is inserted at the Sal I recognition site in pBR322.
Option: 2
In pBR322, when the foreign DNA is inserted at the Hind III recognition site.
Option: 3
When recombinant pBR322 is created by inserting foreign DNA at the Pvu I recognition site.
Option: 4
Recombinant pBR322 that carries foreign DNA at the Pst I recognition site when introduced into a cell.
Answers (1)
- In the pBR322 vector, there are multiple recognition sites for restriction enzymes, including Hind III. These recognition sites are used for inserting foreign DNA during the cloning process.
- When the foreign DNA is inserted at the Hind III recognition site, it disrupts a specific gene responsible for conferring ampicillin resistance in the pBR322 vector. As a result, both recombinant cells (with the foreign DNA inserted at the Hind III site) and non-recombinant cells (without any foreign DNA inserted) would exhibit ampicillin sensitivity.
- Therefore, it becomes difficult to distinguish between recombinant and non-recombinant cells based solely on the ampicillin resistance phenotype. This lack of discrimination makes option (2) the correct answer.
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