Q.Explain the steps involved in the production of recombinant hGH.

Step 1. Isolate the gene: the gene for human growth hormone is isolated from human pituitary gland cells. Step 2. Prepare the vector: a bacterial plasmid is cut open at its sticky ends using the restriction enzyme EcoRI, and the bacterium's own chromosome is left untouched, separate from the plasmid being engineered. Step 3. Build the recombinant DNA: the isolated hGH gene is combined with the cut-open plasmid through DNA recombination, so the plasmid now carries the human hGH gene. Step 4. Transform the host: this recombinant plasmid is introduced into an E. coli cell by DNA insertion, producing a bacterial cell that now contains and can express the gene for human growth hormone. Step 5. Culture and express: the recombinant E. coli are grown in culture; they are then isolated from the culture medium and mass-produced by fermentation technology, during which they manufacture human growth hormone. Step 6. Isolate the product: the recombinant human growth hormone is extracted from the fermentation culture; marketed under the name somatrop …

Walk through Fig. 10.2's pipeline in order: gene isolation from pituitary cells, EcoRI plasmid cutting, recombination, bacterial transformation, f …

• Omitting the specific restriction enzyme (EcoRI) named in the figure and text for opening the plasmid. …