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  • In recombinant DNA technology, which specific enzyme is commonly used to mitigate the issue of unwanted self-ligation of vector DNA molecules?

In recombinant DNA technology, which specific enzyme is commonly used to mitigate the issue of unwanted self-ligation of vector DNA molecules?

Option: 1

DNA polymerase


Option: 2

DNA ligase


Option: 3

Alkaline phosphatase 

 


Option: 4

Reverse transcriptase


Answers (1)

best_answer

In recombinant DNA technology, the enzyme alkaline phosphatase is used to prevent unwanted self-ligation of vector DNA molecules. When a vector, such as a plasmid, is digested with restriction enzymes to create compatible ends for DNA insertion, there is a possibility of the vector molecules re-ligating (joining back together) without the desired insert. To prevent this, alkaline phosphatase is used to dephosphorylate the cut ends of the vector DNA. This dephosphorylation inhibits the self-ligation of the vector, making it unable to recircularize. By preventing self-ligation, alkaline phosphatase increases the efficiency of inserting the desired DNA fragment into the vector during the subsequent ligation step.Hence, the correct answer is option 3.

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