Genetic Engineering and Biotechnology Notes with PDF

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Core Biotechnological Concepts & Processes

Recombinant DNA Technology (Genetic Engineering)

• Core Concept: The technique of manipulating genes by transferring them across different species (e.g., from animals to plants, microorganisms to higher organisms).
  • Scientists can cut and paste DNA molecules from different sources.
  • Functional chromosomes can be created by joining DNA segments from different species.
  • Pieces of artificial functional DNA can be created in laboratories.
  • A piece of DNA from an animal cell can be replicated outside a living cell.
• Key Tools & Enzymes:
  • Restriction Endonucleases: “Molecular Scissors” that cut DNA at specific sequences.
  • DNA Ligases: “Molecular Stitchers” that join DNA fragments together.
  • Cas9 Protein: A type of “molecular scissors” used in targeted gene editing.
• Process Steps (Sequence):
  1. Identification and isolation of the genetic material.
  2. Fragmentation of DNA.
  3. Isolation of desired DNA fragments.
  4. Amplification of the gene of interest.
  5. Ligation of the DNA fragment into a vector.
  6. Transfer of the Recombinant DNA into a host.
  7. Obtaining the foreign gene product.
  8. Downstream processing.
• Applications: Vaccines
  • Recombinant Vector Vaccines are developed using genetic engineering, using bacteria and viruses as vectors.

RNA Interference (RNAi) Technology

• Applications & Significance:
  • Developing gene silencing therapies.
  • Therapies for the treatment of cancer.
  • Producing crop plants resistant to viral pathogens.

Cloning

• General Principles:
  • A clone is produced asexually.
  • Identical twins are a natural example of clones.
  • Animal cloning is a successful application of genetic engineering.
• Landmark Clones:
  • Dolly the Sheep: First mammal cloned from an adult somatic cell.
    • Born in 1996 in Scotland.
    • Died in 2003 due to a lung disease.
    • Produced by nuclear transfer, not by in-vitro fertilization.
  • Other Clones: Garima-II (a cloned buffalo in India), Injaz (world’s first cloned camel).
  • The first successful cloned animal was a sheep (Dolly).

Transgenic Organisms

• Glowing Pigs: The first transgenic glowing pigs (green inside out) were produced in Taiwan.

Human Applications & Reproductive Technologies

Human Reproductive Technologies

• In-Vitro Fertilization (IVF) / Test-Tube Baby:
  • Fertilization takes place outside the mother’s body (e.g., in a petri dish).
  • The resulting embryo is then implanted.
• Mitochondrial Replacement Therapy:
  • Pronuclear Transfer is a technique used for it.
  • Its purpose is the prevention of mitochondrial diseases in offspring.
  • This therapy is performed before in vitro fertilization of the egg.
  • A child inherits mitochondrial DNA entirely from the mother.

Stem Cell Research

• Definition & Potential:
  • Stem cells are undifferentiated cells with the capacity to divide and differentiate into any type of cell.
  • Focus of research for serious diseases due to their ability to repair damaged tissues.
  • Can be used for medical therapies and for screening new drugs.
• Sources and Properties:
  • Sources are not limited to mammals or embryos.
  • Can be derived without destroying blastocysts (e.g., from adult tissues).
  • A bioethically non-controversial source is bone marrow-derived stem cells.
  • They can regenerate themselves in vitro for a very long time.
  • They are not found only in embryos; adult stem cells exist.
• Indian Contribution: Indian research centers have created cell lines that can be developed into many tissue types.

Agricultural Biotechnology & GM Crops

Genetically Modified (GM) Crops: Concepts & Objectives

• Objectives of Genetically Engineered Plants:
  • Drought tolerance
  • Enhanced nutrition (e.g., Golden Rice)
  • Extended shelf life (e.g., Flavr-Savr Tomato)
  • Pest resistance (e.g., Bt crops)
• Techniques for Creating Transgenic Crops:
  • Gene Silencing: Used to alter traits by switching off specific genes.
  • Cytoplasmic Male Sterility: Used in plant breeding, particularly for developing hybrids.
  • (Budding and grafting are conventional techniques and not used for transgenics).

Specific GM Crop Examples

• Bt Technology:
  • Bt stands for Bacillus thuringiensis, a bacterium.
  • The Bt toxin produced by the plant is lethal to specific insect pests.
  • Bt Cotton: A prime example; includes technologies like Bollgard I and Bollgard II.
  • Bt Brinjal: A genetically modified brinjal developed to be pest-resistant.
• GM Mustard (India):
  • Developed to contain genes that facilitate cross-pollination and hybridization.
  • Developed by Indian agricultural institutions (e.g., IARI).
• Golden Rice:
  • A transgenic plant developed to combat Vitamin A deficiency.
  • The rice kernel is rich in Beta-Carotene (converted to Vitamin A), giving it a golden color.
  • The β-carotene gene was inserted from a daffodil plant.
• Flavr-Savr Tomato:
  • The first genetically engineered crop product to be commercialized.
  • Engineered for a longer shelf life; ripe fruits remain firm longer.

Terminator Seed Technology

• Officially known as Gene Use Restriction Technology.
• It produces genetically engineered seeds that are sterile in the next generation.
• This prevents farmers from saving seeds, forcing them to buy new seeds each season.

Supporting Technologies & Concepts

Bio-pesticides & Microbial Insecticides

• Species of Bacteria, Fungi, and Flowering plants can be employed as bio-pesticides.
• Bacillus thuringiensis (Bt) is a species of bacteria used as a microbial insecticide.

Genome Sequencing in Agriculture

• Used to identify genetic markers for traits like disease resistance.
• Helps in reducing the time to develop new crop varieties.
• Can be used to decipher host-pathogen relationships.

Cell Biology Fundamentals

• Cells taken from plants and animals can be made to undergo cell division in laboratory petri dishes.

Genetic Analysis, Forensics, and Diagnostics

DNA Fingerprinting & Forensic Science

• Principle: Based on DNA polymorphism (variations in DNA sequences).
• Applications:
  • Establishing paternity and identity.
  • Solving forensic cases (e.g., rape, assault).
  • Conservation of endangered species.
• History: The first crime solved using DNA fingerprinting was in England in 1987.
• Biometric Identification can also use Iris scanning, Retinal scanning, and Voice recognition.

Basic Genetics & Medical Diagnostics

• Genetic Phenomena:
  • Mutation: A change in the base sequence within a gene.
  • Pleiotropy: When a single gene controls two or more different characteristics.
• Medical Techniques:
  • Amniocentesis: Can be used to determine foetal sex.
  • Gene Therapy: Aims to correct a faulty gene by inserting a functional gene.

Key Scientists, Projects, and Rights

Key Scientists and Discoveries

• Joshua Lederberg: Discovered transduction and conjugation in bacteria.
• Thomas Hunt Morgan: Established principles of sex-linked inheritance.
• Arthur Kornberg: Isolated DNA polymerase from E. coli; achieved first in-vitro DNA synthesis.
• Har Gobind Khorana & Severo Ochoa: Key roles in establishing the complete genetic code.

The Human Genome Project (HGP)

• A world-level program to identify and map all human genes and their sequences.
• Benefits: Allows for understanding livestock pedigrees and developing disease-resistant animal breeds.
• Limitation: It is not possible to understand the causes of all human diseases from it.
• First Sequenced Organism: The genome of Yeast (Saccharomyces cerevisiae) was fully sequenced in 1996.

Biodiversity and Intellectual Property Rights (IPR)

• Plant Field Gene Bank (e.g., at Banthara) serves to:
  • Preserve endangered plant varieties.
  • Check piracy of biodiversity.
  • Identify economically important wild plants.
• Indian Patents Act:
  • In India, plant varieties are not eligible to be patented.
  • A biological process to create a seed (like conventional breeding) cannot be patented.
  • The Intellectual Property Appellate Board (IPAB) has been dissolved.

Advanced Genetic Techniques & Definitions

• Biotechnology: The use of organisms or their substances in industrial processes.
• Gene Splicing: A recent evolution in genetic engineering techniques.
• Key Tools: Plasmids (small, circular DNA molecules) and Bacteriophages (viruses that infect bacteria) are used as tools.
• Transgenics: Involves gene transfer to produce transgenic crops, edible vaccines, etc. (Note: Cloning is a separate technique).
• Genico Technology: A technique for obtaining pre-information regarding genetic diseases.
• Transcriptome: The full range of mRNA molecules expressed by an organism.
• Biochip: Typically contains RNA, DNA, and Protein.
• Advanced Techniques:
  • Introducing genetic changes in germ cells (egg/sperm).
  • Editing a genome at the early embryonic stage.
  • Injecting human stem cells into the embryo of another animal (e.g., a pig).
  • Somatic Cell Nuclear Transfer (SCNT): Primary application is the reproductive cloning of animals.
  • Hybridoma Technology: Used for the commercial production of monoclonal antibodies.

Know More About Genetic Engineering and Biotechnology:

• Genetic Engineering and Biotechnology – Old Year Questions
• Genetic Engineering and Biotechnology One Liner Questions & Answers