Watson and Crick Model

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Biotechnology In Local Schools (BILS)

Presented by
Holly Jefferson Biotechnology Instructor hjefferso
n_at_lenoircc.edu And Mickey Adams Biology
Instructor madams_at_lenoircc.edu
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Introduction to BILS

• Provides an opportunity for teachers and
  students to learn the fundamentals of
  biotechnology, genetics, and/or molecular
  biology.
• Introduces career paths in the field of
  biotechnology to students.
• Offers dual enrollment to high school students
  to Lenoir Community College.
• Demonstrations and hands on activity in local
  middle and high schools performed by LCC staff
  with the use of mobile labs.
• BILS manual to serve as a resource for middle
  and high school teachers.

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What is Biotechnology?

• Biological science when applied especially in
  genetic engineering and recombinant DNA
  technology - Meriam-Webster

The use living organisms or their
products to perform a valuable purpose.
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History of Biotechnology

• 8000 BC Mesopotamia Humans began farming and
  the domestication of animals.

Selective breeding soon followed.
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History of Biotechnology

• 500 BCIn China, moldy soybean curds became first
  antibiotic used to treat infections/ailments
• 300 BCGreeks develop grafting techniques for
  plant breeding.
• 100 ADFirst insecticide produced in China from
  powdered chrysanthemums.

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History of Biotechnology

• 1663 Robert Hooke discovers Cells
• 1861 Louis Pasteur develops pasteurization and
  Germ Theory
• 1865 Gregory Mendel, the father of modern
  genetics, discovers laws of heredity

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History of Biotechnology

• Late 1800s
• Luther Burbank, the father of modern plant
  breeding, developed more than 800 new strains of
  fruits, vegetables and flowers.
• Plant breeders crossbreed cotton to develop
  hundreds of varieties with superior growing
  qualities.
• Farmers first inoculate fields with
  nitrogen-fixing bacteria to improve yields.
• First experimental corn hybrid produced in the
  laboratory by William James Beal, a Professor of
  Botany, at Michigan Academy of Sciences,
  1870-1910.

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History of Biotechnology

• 1941 Danish microbiologist A. Justin coins term
  "genetic engineering
• Technique involving a transfer of a select piece
  of genetic material from one organism to another.
  
• Problem We did not yet know what the genetic
  material was.

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History of Biotechnology

• In 1944 Oswald Avery proves DNA is the genetic
  material, and his findings were confirmed in 1952
  by Hershey and Chase.

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In 1953, DNA (deoxyribonucleic acid) was
discovered as a double helix structure by Watson
and Crick using data collected by Rosalind
Franklin.
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History of Biotechnology

• 1961 Marshall Nirenberg and colleagues decipher
  the genetic code
• Genetic code is a triplet code, with each codon
  consisting of three nucleotide bases.
• Code Properties
• Universal
• Degenerate
• Unambiguous
• Contains start and stop signals
• Common Origin of Life

Marshall Nirenberg
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History of Biotechnology

• 1970 American microbiologist, Daniel Nathans,
  discovers first restriction enzyme which can cut
  specifically cut DNA at a specific recognition
  site.

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History of Biotechnology

• 1972 DNA ligase, which links DNA fragments
  together, used for the first time.
• 1973 Stanley Cohen and Herbert Boyer discover
  recombinant DNA technology. They inserted a gene
  from an African clawed toad into bacterial DNA.
• 1978 Boyer was the first to insert a human gene
  (Insulin) into Bacteria for the first time.
• 1982 Scientists at Ohio University produced the
  first transgenic animals by transferring genes
  from other animals into mice.

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History of Biotechnology

• In 1983 the first genetically engineered drug was
  developed.
• Genentech's human insulin drug produced by
  genetically engineered bacteria was the first
  biotech drug to be approved by the Food and Drug
  Administration for the treatment of Diabetes.

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History of Biotechnology

• 1985 First genetically modified crops field
  tested.
• 1986The EPA approved the release of the first
  genetically engineered crop, virus resistant
  tobacco plants.
• 1990 Human Genome Project launched. Aim
  Sequence and Map the entire genome.
• 1994 The first genetically engineered food
  product, the Flavr Savr tomato, gained FDA
  approval
• 2003 Human Genome successfully mapped.

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Biotechnology Today

• Agricultural Biotechnology
• Genetically altered Crops
• Genetically modified Animals
• Environmental Biotechnology
• Bioremediation
• Environmental Testing
• Energy Procurement
• Materials Science

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Biotechnology Today

• Medical Biotechnology
• Medical Research
• Disease Identification
• Genetic Screening
• Designer Drugs/Vaccines
• Gene Therapy
• Tissue/Organ Engineering
• Forensics
• DNA Fingerprinting

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How big is Biotech?

• 450 Billion dollars in annual revenues with
  projected annual growth of 10-15.
• The US accounts for 70 of the Global Biotech
  Industry.
• Biomanufacturing jobs are among the highest
  paying in the manufacturing sector.
• Starting wages of 25,000-30,000 per year that
  can grow to 40,000-50,000 with experience.

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NC Biotech
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Importance of Biotech to NC

• Wetlands bioremediation or aquaculture
• Plant and Animal agriculture tobacco and sweet
  potatoes 1 crops, hogs and broilers 1 farm
  commodities
• Forest Industry christmas trees 2 crop
• Medical Research

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Importance of Biotech to NC

• NC has the 3rd largest biotech industry in the
  nation employing 48,897 people with an annual
  payroll of 1 Billion, and generates 3
  Billion in revenue per year.
• Milken Institute report predicts 7,000 new
  biotech jobs in NC by 2014.
• 67 of workers have HS diploma, certification, or
  AAS, 27 have BS, and only 6 have post graduate
  degrees.
• Community Colleges play a vital role in training
  biomanufacturing workers.

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Classroom Lecture Material

• Objective
• To understand the structure and function of
  cells, understand the importance of DNA and RNA,
  and to understand the basic principles of
  Biotechnology
• Classroom Lecture contains
• Cell structure and function
• DNA and RNA structure and function
• Recombinant DNA technology
• Gel electrophoresis

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The Cell

• The cell is the building block of all living
  things
• All organisms are composed of Cells
• Cells can only come from preexisting cells
• There are two general types of cells
• Prokaryotic Cells like bacterial cells are simple
  cells that lack a nucleus and membrane bound
  organelles
• Eukaryotic cells are much more complex, contain a
  nucleus and membrane bound organelles

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The Cell

• Cell Membrane
• Separates cells from the environment
• Regulates permeability of ions, nutrients, and
  waste products
• Composed of a Phospholipid bilayer
• Cytoplasm
• Contains Cytosol, the watery liquid of the cell
  and cellular organelles
• Organnelles are structures that perform specific
  functions inside cells

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The Cell

• Cytoskeleton
• Serves and skeleton for the cells providing
  support, strength, stability, and plays a role in
  transport of cellular products
• Microvilli
• Increase the surface areas of the cell to allow
  for increased absorptive capacity.
• Centrosome
• Assists in movement of chromosomes during
  cellular division

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The Cell

• Cilia
• Slender extensions of the cell membrane that beat
  rhythmically to move fluids across the surface of
  cells or provide locomotion
• Ribosomes
• Organelles responsible for the translation of
  mRNA into protein
• Proteasome
• Organelles responsible to breaking down cellular
  proteins

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The Cell

• Endoplasmic Reticulum
• A network of intracellular membranes connected to
  the nuclear envelope surrounding the nucleus of a
  cell
• Synthesis, storage, transport, and detoxification
  
• Golgi Apparatus
• Modifyies and packages secretions, such as,
  enzymes, for release through exocytosis, it
  renews or modifies the cell membrane, and
  packages special enzymes within vesicles for use
  in the cytosol of the cell

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The Cell

• Lysosomes
• Organelles that contain digestive enzymes that
  function in the breakdown of foreign and
  intracellular materials
• Peroxisome
• Organelles that break down fatty acids and other
  organic compounds

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The Cell

• Nucleus
• The nucleus serves as the headquarters of
  cellular activities
• Stores and processes DNA

• Mitochondria
• The organelle is the power plant of the cell.
  The mitochondria produces energy for cellular
  activities in the form of ATP

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DNA

• Double stranded polymer that stores an organisms
  genetic information
• Composed of phosphoric acid, a pentose sugar
  called ribose, and a nitrogenous base
• Two strands of nucleotides intertwine to form a
  double helix structure

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DNA

• The nitrogenous bases exhibit complimentary base
  pairing and form hydrogen bonds that hold the two
  strands of DNA together
• The pyrimidines cytosine, and thymine for
  hydrogen bonds with the purines guanine and
  adenine respectively

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The Genetic Code

• An organisms genetic information, the organisms
  genes, is written in a DNA language.
• A gene is a segment of DNA that codes for a
  particular polypeptide product. These
  polypeptides called proteins perform numerous
  critical roles in each cell in an organism.
• The sequence of nitrogenous bases of the DNA
  molecule contains a code that determines the
  particular amino acids that will be incorporated
  into the protein.

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The Genetic Code

• Each protein is composed of different combination
  of 20 different amino acids
• Three nitrogenous bases code for one amino acid
  in a protein, therefore the genetic code is a
  triplet code
• One triplet is called a codon
• Code properties
• The code is universal for all organisms
• More than one codon codes for the same amino acid

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How Genes Work

• DNA is Transcribed into mRNA and mRNA is
  Translated into Protein
• RNA is a very similar to DNA with a few
  structural differences

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Recombinant DNA Technology

• Several discoveries combined with the knowledge
  of DNA structure and function allowed scientists
  to construct DNA molecules in the test tube.
• These recombinant DNA molecules can be placed in
  other organisms in order to manufacture a protein
  product of interest or make copies of the gene.
  This is commonly referred to a gene cloning.
• The process of placing foreign DNA into a host
  organism is called transformation.
• Organisms created in this way are referred to as
  transgenic.
• Many beneficial molecules can be manufactured
  this way including insulin, growth hormone, and
  antibodies.

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Recombinant DNA Technology

• The Recombinant DNA Technology Toolkit
• Restriction enzymes specifically cleave DNA and
  thus act as DNA scissors
• DNA ligase An enzyme that binds to two DNA
  fragments together
• Vector Serves to carry foreign DNA inside host
  cells.
• Plasmids Circular strands of bacterial DNA that
  can easily transport foreign DNA inside cells
• Viruses Intracellular parasites that can be
  harnessed to transport foreign DNA inside cells

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Recombinant DNA Technology
How to

• Isolate gene of interest
• Ligate gene into a vector
• Transform host organism
• Culture host organism
• Purify gene product of interest

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Production of Transgenic Organisms

• The production of transgenic organisms involves
  the injection of foreign DNA into an egg.
• The egg is then fertilized and placed inside a
  surrogate organism which carries the transgenic
  organism to term.

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Production of Transgenic Plants

• Transgenic plants can be produced with plant cell
  culture.
• Foreign DNA is used to transform disassociated
  plant cells that are then grown in culture.

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Gene Therapy

• Researchers are currently working on ways to
  treat and even cure certain genetic disorders
  utilizing recombinant DNA technology
• Today many can be detected using genetic screening

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DNA Gel Electrophoresis

• Fragments of DNA can be separated based on size
  when subjected to an electrical field because DNA
  carries a negative charge.
• DNA samples are loaded into semisolid matrix made
  of the carbohydrate agarose.
• As the DNA moves through the gel the DNA collides
  with the matrix. The smaller fragments collide
  less often and thus move faster than larger
  fragments
• DNA Gel electrophoresis allows DNA to be
  visualized after staining

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Gel Electrophoresis
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Uses of DNA Electrophoresis

• Identify people by their DNA (DNA fingerprinting)
• Test for diseases
• Allows manipulation of fragments
• Gene mapping

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DNA Fingerprinting

• Everyone's DNA has small differences. The only
  exceptions are identical siblings.
• When subjected to restriction enzymes that cut
  specific sequences of DNA, each persons DNA will
  be cut into different sized fragments. These are
  referred to as restriction fragment lengths
  polymorphisms or RFLPs
• Since each person has different RFLPs, each
  individuals fragments produce different banding
  patterns when subjected to gel electrophoresis.
  The result is a unique pattern of DNA fragments
  called a DNA fingerprint.

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Classroom Activities

• The classroom activities detailed in the BILS
  manual are intended to foster understanding of
  cell biology and biotechnology.
• Activities described in the manual are
• Building a cell Understand the structure of the
  cell by building a 3-D model
• Playing a cell Understand basic cell physiology
  and organelle function by acting out the
  structures of the cell
• Diffusion and Osmosis Explain the concepts of
  diffusion and osmosis visually
• Micropipetting Introduce the students to the
  most common tool used in biotechnology
  laboratories and also review the metric system

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Micropipetting
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Classroom Activities

• Activities described in the manual continued
• DNA Isolation Students isolate and can actually
  see DNA from strawberries or kiwi fruit using
  common household items
• DNA Gel Electrophoresis Students learn to
  perform one of the most fundamental techniques of
  biotechnology whereby DNA is separated in an
  agarose gel matrix according to size

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For More Information

• For more information on BILS www.lenoircc.edu/bio
  tech/
• For the Biotechnology Workshop Manual
  www.lenoircc.edu/biotech/BiotechWorkshop.pdf
• Contacts
• Holly Jefferson hjefferson_at_lenoircc.edu
• Lisa Boyd lboyd_at_lenoircc.edu

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Acknowledgements

• Lisa Boyd
• Maria Pharr