Biotechnology Principles and Processes

Introduction

• Lecture by Dikshama on biotechnology principles and processes.
• Important for CBSE and NEET exams.
• Biotechnology is an emerging field in science.
• Opportunities in biotechnology for students not wanting to pursue medicine.

What is Biotechnology?

• Definition by the European Federation of Biotechnology:
  • Integration of natural science, organisms, and molecular analogs for products and services.
• Combination of Biology and Technology (Engineering).
• Use of natural organisms, cells, or molecular analogs to create products/services.
• Example: Production of insulin in labs as a molecular analog.

Types of Biotechnology

1. Traditional Biotechnology
   • Historical applications (e.g. curd, fermented batter).
2. Modern Biotechnology
   • Advanced techniques and controlled environments (e.g. large-scale production of curd in industries).

Principles of Biotechnology

• Focus on techniques and processes:
  • Techniques: Methods used in biotechnology.
  • Processes: Application of these techniques.
• Two basic principles:
  1. Genetic Engineering
     • Involves cutting and joining DNA (forming recombinant DNA).
     • Introduction of DNA into a host to produce desired products.
  2. Maintenance of Sterile Culture
     • Ensuring contamination-free conditions during experiments.

Genetic Engineering Process

• Gene Cloning:
  • Isolate a gene of interest (e.g., insulin gene from beta cells).
  • Use Restriction Enzymes to cut DNA at specific sites (e.g., EcoRI).
  • Use Vectors to transport the gene into a host (e.g., E. coli).
  • Process called Transformation when recombinant DNA is introduced into the host.

Important Tools in Recombinant DNA Technology

1. Restriction Enzymes:
   • Molecular scissors that cut DNA at specific sequences.
   • Types include sticky ends and blunt ends based on cut style.
2. Ligases:
   • Join DNA fragments by forming phosphodiester bonds.
3. DNA Polymerase:
   • Replicates DNA.
4. Alkaline Phosphatase:
   • Cleaves phosphate groups to prevent rejoining.
5. Vectors:
   • Transport the gene of interest into host cells.
   • Must have origin of replication, selectable markers, and multiple cloning sites (MCS).
   • Example: pBR322 (contains ampicillin and tetracycline resistance genes).

Vector Types

1. Plasmids: Circular DNA in bacteria.
2. Bacteriophages: Viruses that infect bacteria (e.g. lambda phage).
3. Cosmids: Hybrid vectors.
4. YACs and BACs: Yeast Artificial Chromosomes and Bacterial Artificial Chromosomes for larger inserts.
5. Ti Plasmid: Used in plants (e.g., Agrobacterium tumefaciens).
6. Retroviruses: Used in gene therapy to deliver genes into animals.

Processes in Biotechnology

1. Isolation of Genetic Material: Extracting DNA from cells.
2. Cutting DNA: Using restriction enzymes to create fragments.
3. Separation and Isolation: Using gel electrophoresis to visualize DNA fragments.
4. Amplification: Polymerase Chain Reaction (PCR) to replicate DNA.
5. Transformation: Introducing the recombinant DNA into host cells.
6. Culturing: Growing transformed cells in bioreactors to produce the desired product.
7. Downstream Processing: Isolating and purifying the product.

Summary of Techniques

• Transformation Techniques:
  • Calcium chloride treatment, heat shock, microinjection, biolistics (gene gun), electroporation.
• Selection Techniques:
  • Blue/White selection and Insertional Inactivation to identify recombinant DNA.

Bioreactors

• Large vessels designed for culturing microorganisms.
• Types: Batch reactors (fixed quantity) and Continuous reactors (constant flow).
• Equipment includes sterilization steam, stirrers, and pH control.

Conclusion

• Biotechnology is a complex yet fascinating field with practical applications in medicine, agriculture, and industry.
• Continuous learning and hands-on practice are essential for mastery.