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How are plasmids used as vectors in recombinant DNA technology?
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Nucleic acid
A portion of the polynucleotide chain of deoxyribonucleic acid (DNA), with an inset showing the corresponding pentose sugar and pyrimidine base in ribonucleic acid (RNA).
Plasmids are circular, extrachromosomal deoxyribonucleic acid (DNA) molecules that serve as essential vectors in recombinant DNA technology by acting as carriers for foreign genetic material.12
The process of using plasmids as vectors involves several key steps:
- Preparation: A plasmid is cut at specific sites using restriction enzymes, which function as molecular scissors to create openings for foreign DNA.13
- Splicing: A foreign DNA fragment is inserted into the opened plasmid. Because the donor DNA and the vector are often cut with the same enzyme, they possess complementary single-stranded overhangs that allow them to pair together.3
- Ligation: An enzyme called DNA ligase is applied to seal the sugar-phosphate backbone, creating a stable, continuous recombinant DNA molecule.3
- Transformation and Amplification: The recombinant plasmid is introduced into bacterial cells through transformation. Once inside, the plasmid replicates autonomously as the bacteria divide, producing large numbers of copies of the inserted DNA.13
Plasmids are ideal for this purpose because they are small, easily manipulated, and can carry extra DNA sequences for applications like gene cloning and protein production.12