How do you insert a gene into a plasmid?

Question

Ruby Armstrong · Accepted Answer

A gene is nothing but a portion of dna coding for a specific protein.

A DNA insert is simply the gene that is going to be added to the plasmid. Even the circular plasmid is cleaved using the same or different restriction enzyme based on the requirement. (the cleaving of the circular plasmid is to create space to insert the DNA insert). The gene of interest or the DNA insert is initially cleaved from the genomic source of an organism using an enzyme known as the restriction endonuclease or is artificially synthesized in the laboratory. Now, a powerful gluing enzyme called DNA ligase is used to attach the DNA insert to the plasmid (T4 ligase from bacteriophage is the type of DNA ligase enzyme that is usually used). Additionally, the entire recombinant vector—that is, the plasmid plus DNA insert—is built.

Piper Anderson · Answer

undefined To insert a gene into a plasmid, you can use a technique called molecular cloning. This involves cutting both the plasmid DNA and the gene of interest with restriction enzymes, which produce compatible ends. Then, the gene is ligated into the plasmid using DNA ligase. This results in a recombinant DNA molecule containing the gene of interest inserted into the plasmid.