Genetics: Genetic Technology

Unit 5: Genetic Technology

• Genetic technology involves the manipulation and analysis of DNA at the molecular level.
• Recombinant DNA technology is used to alter genetic material outside an organism to obtain desired characteristics.
• This technology involves the insertion of DNA fragments from various sources into a vector, using appropriate enzymes and procedures.

Recombinant DNA Technology

• The process involves:
  • Isolation of genetic material
  • Restriction enzyme digestion
  • Amplification using PCR
  • Ligation of DNA molecules
  • Insertion of recombinant DNA into a host
• Recombinant DNA technology has advanced strategies for biomedical applications, including cancer treatment, genetic diseases, and diabetes.

Gene Cloning

• Gene cloning involves making multiple copies of a gene.
• The process involves:
  • Isolation of chromosomal DNA
  • Digestion with restriction enzymes
  • Ligation of chromosomal DNA into a vector
  • Transformation of host cells
  • Selection of recombinant cells
• Gene cloning has provided the foundation for critical technical advances in various disciplines, including molecular biology, genetics, and medicine.

The Restriction Enzyme

• Restriction enzymes are used to cut DNA at specific locations.
• They are named according to the species in which they are found.
• Examples of restriction enzymes include EcoRI, PstI, and NaeI.

PCR (Polymerase Chain Reaction)

• PCR is a method of making multiple copies of a DNA sequence.
• The process involves:
  • Denaturation
  • Primer annealing
  • Primer extension
• PCR is carried out in a thermocycler, which automates the timing of the temperature changes.
• The advantages of PCR include:
  • Amplification of a specific region of DNA
  • Ability to amplify DNA from a complex mixture of templates

Biotechnology

• Biotechnology is the use of living organisms or substances they produce in the development of products or processes that are beneficial to humans.
• Molecular genetic tools have provided novel ways to make use of living organisms.
• Recombinant methods enable the introduction of genetic material into animals and plants, resulting in genetically modified organisms (GMOs).

Microorganism in Biotechnology

• Microorganisms are used to produce various medical agents, including:
  • Insulin
  • Tissue Plasminogen Activator (TPA)
  • Superoxide dismutase
  • Factor VIII
  • Renin inhibitor
  • Erythropoietin### Biotechnology Applications
• Erythropoietin (EPO) stimulates the production of red blood cells, used to treat anemia.
• Food Fermentation: utilizes microorganisms to stabilize and transform food materials, examples include cheese, yogurt, vinegar, and wine.

Biological Control

• Biological control: uses living organisms or their products to alleviate plant diseases or environmental conditions.
• Bacillus thuringiensis (Bt): a bacterium producing toxins lethal to caterpillars and beetles that feed on crops.
• Bt is harmless to plants and beneficial insects, making it an environmentally friendly pesticide.

Bioremediation

• Bioremediation: uses living organisms or their products to decrease pollutants in the environment.
• Microorganisms modify toxic pollutants through biotransformation, resulting in their degradation into non-toxic metabolites.

Genetically Modified Animals

• Genetically modified animals: altered to introduce a new gene or modify an existing gene.
• Methods include gene modification (altering the sequence of a gene) and gene addition (introducing a cloned gene into a genome).
• Example: GloFish, a genetically modified aquarium fish that glows due to a fluorescent protein gene from jellyfish or sea coral.

Reproductive Cloning and Stem Cells

• Reproductive cloning: produces genetically identical individuals, naturally occurring in identical twins.
• Researchers can clone mammals from somatic cells, as evidenced by Dolly the sheep.
• Stem cells: supply various cell types in the body, can differentiate into multiple cell types.
• Stem cells have the capacity to divide and differentiate into specialized cells.

Human Gene Therapy

• Gene therapy: introduces cloned genes into somatic cells or modifies existing genes to treat diseases.
• Targets diseases such as cancer and cardiovascular disease.
• Some results have been promising, but success has been limited.

Genomics

• Genomics: analyzes the entire genome of a species.
• Genome: the total genetic composition of an organism or species.
• Genome analysis is a molecular dissection process applied to a complete set of chromosomes.

Chromosome Mapping

• Genetic map: a diagram describing the relative locations of genes or DNA segments along a chromosome.
• Cytogenic mapping: determines the locations of specific sequences within chromosomes using microscopy and staining techniques.
• Linkage mapping: uses the frequency of recombinant offspring to determine the distance between sites along a chromosome.
• Physical mapping: establishes a physical map of a genome through cloning and DNA sequencing.

Metagenomics

• Metagenomics: studies complex mixtures of genetic material from environmental samples.
• Applications:
  • Human medicine: characterizes microorganisms associated with diseases.
  • Agriculture: identifies microorganisms that facilitate plant growth.
  • Bioremediation: identifies microorganisms that break down pollutants.
  • Biotechnology: discovers genes encoding useful products.
  • Global change: understands the complexity of microbial communities.
  • Identification of viruses: analyzes environmental samples to identify viruses.

Functional Genomics, Proteomics, and Bioinformatics

• Functional genomics: aims to understand the roles of genetic sequences in a species.
• Methods:
  • DNA microarrays: quantify the expression of thousands of genes simultaneously.
  • RNA sequencing: sequences complementary DNAs derived from RNAs.
• Proteomics: identifies and quantifies proteins in a cell, tissue, or organism.
• Bioinformatics: combines mathematics, information science, and biology to answer biological questions.### Bioinformatics Components
• Bioinformatics consists of two main components:
  • Developing software tools and algorithms
  • Analyzing and interpreting biological data using software tools and algorithms

Biotechnology in Daily Life

• Biotechnology is used in various products and processes that we use daily
• Examples of biotechnology-related products/processes and their importance to our wellbeing include:

Biotechnology Examples

• Product/Process: Food Fermentation
  • Specific Biotechnology: Microbial fermentation
  • Species Involved: Microorganisms (e.g., yeast, bacteria)
  • Importance to Wellbeing: Provides nutritious and safe food products (e.g., bread, yogurt, cheese)

References

• Important sources for genetics and biotechnology include:
  • Brooker, R.J. (2018). Genetics: Analysis and Principles (6th Edition)
  • Reece, J.B., et al. (2010). Campbell Biology (10th Ed)
  • Shiferaw Terefe, N. (2016). Food Fermentation
  • Aslam, B., et al. (2017). Proteomics: Technologies and Their Applications