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Question

A recombinant DNA molecule was created by ligating a gene into a plasmid. By mistake, an exonuclease was added to the tube containing the rDNA. How will this affect the further steps of this technology?


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Solution

Recombinant DNA technology:

  1. Recombinant DNA molecule involves the process of joining two or more different DNA molecules of interest from different species, insertion, and its expression into a suitable host.
  2. It is created by ligating the ‘gene of interest’ into a plasmid (vector) with the help of DNA ligase.
  3. Plasmids are extrachromosomal small DNA naturally found in bacteria, that self-replicates irrespective of genomic DNA replication.
  4. An exonuclease is a type of restriction enzyme that remove nucleotides from the free end of a DNA molecule. They act as proofreaders during DNA polymerization to remove any unusual fragments.
  5. Example: Exonuclease I, Xrn1.
  6. Adding an exonuclease to the tube containing the rDNA will not affect further steps of this technology. Recombinant DNA is circular with no free ends; hence exonuclease cannot cleave this kind of DNA.

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