Cloning DNA using plasmids includes 3 steps.
1. Cutting and pasting DNA – A restriction enzyme recognises a particular target sequence and cuts DNA into two halves. Several restriction enzymes generate cut ends with short and single-stranded overhangs. However, if two molecules have similar overhangs, then they can base-pair and stick together.
2. Bacterial selection and transformation – Plasmids can be induced into bacteria through a process called transformation. During transformation, the prepared bacterial cells are given a shock thereby encouraging them to take up foreign DNA. A plasmid consists of an antibiotic resistance gene that enables bacteria to survive even in the presence of a specific antibiotic. Thus, bacteria that takes the plasmid can be selected.
3. Protein production – Once the bacterial colony is founded with the right plasmid, you can grow a large culture of plasmid-bearing bacteria. Also, the bacteria serve as factories churning a huge amount of protein.
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