Revision: Biotechnology and Its Applications >> Biotechnology - Principles and Processes CUET (UG) Biotechnology - Principles and Processes

The technique of bringing about improvements in living organisms by genetic modifications and hybridization, for the welfare of human beings is known as ‘Biotechnology’.

The European Federation of Biotechnology (EFB) defined biotechnology as ‘the integration of natural science and organisms, cells, parts thereof, and molecular analogues for products and services.’

• Biotechnology involves the use of biological systems, cells, and biomolecules along with genetic and non-genetic techniques for human welfare.
• It integrates sciences such as cytology, biochemistry, molecular biology, and genetic engineering and has major applications in agriculture and pharmacy.
• Biotechnology enables production of antibiotics, vaccines, hormones (insulin), antibodies, and development of high-yield crop varieties through tissue culture and gene manipulation.
• Major benefits include increased crop yield, development of disease- and stress-resistant varieties, and reduced expenditure on disease control.
• In India, biotechnology is promoted by the Department of Biotechnology (DBT) established in 1986, under which several national research institutes function.

• Recombinant DNA technology depends on the isolation, cutting, and joining of DNA fragments to form chimeric DNA.
• Restriction enzymes act as molecular scissors that cut DNA at specific sequences to obtain desired genes.
• DNA ligase joins DNA fragments by forming covalent bonds, producing recombinant DNA molecules.
• Cloning vectors are DNA molecules that carry foreign DNA into host cells for replication.
• Common vectors include plasmids, bacteriophages, cosmids, and phasmids, which replicate independently inside host cells.

1. Isolation of DNA:
   Genetic material (DNA) is extracted and purified from the source organism by removing proteins, RNA, lipids, and other impurities.
2. Cutting and Separation of DNA:
   DNA is cut at specific sites using restriction endonucleases, and desired fragments are separated by gel electrophoresis and isolated by elution.
3. Amplification of Gene (PCR):
   The gene of interest is amplified using Polymerase Chain Reaction (PCR) to produce millions of copies for effective cloning.
4. Ligation and Gene Transfer:
   The desired DNA fragment is ligated into a vector and introduced into a host cell by methods such as transformation, electroporation, or gene gun.
5. Expression and Production:
   The host cells expressing the recombinant gene are cultured on a large scale in bioreactors to produce the desired recombinant protein.
6. Downstream Processing:
   The final product is separated, purified, and processed to obtain a market-ready product.