Question

Give the steps involved in construction of recombinant DNA molecule.

Answer 1

DNA cloning is used for many purposes. As an example, let's see how DNA cloning can be used to synthesize a protein (such as human insulin) in bacteria. The basic steps are:

• Cut open the plasmid and "paste" in the gene. This process relies on restriction enzymes (which cut DNA) and DNA ligase (which joins DNA).
• Insert the plasmid into bacteria. Use antibiotic selection to identify the bacteria that took up the plasmid.
• Grow up lots of plasmid-carrying bacteria and use them as "factories" to make the protein. Harvest the protein from the bacteria and purify it.