Today

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Today s Agenda

• Journal Questions
• (1) Describe what you know about DNA.
• (2) What is genetic engineering (Biotechnology)?
• 1. Lecture Genetic Engineering (Biotechnology)
  Recombinant DNA Technology -slide . 80

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Genetically Modified (GM) Crops around the World
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Genetic Engineering

• Dr. Rick Woodward

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Genetic Engineering
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Genetic Engineering
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Genetically Engineered Boneless Chicken Ranch
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DNA, the Law, and Many Other Applications The
Technology of DNA Fingerprinting
A DNA fingerprint used in a murder case.
The defendant stated that the blood on his
clothing was not his.
What are we looking at? How was it produced?
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DNA Fingerprinting Basics
A. Different individuals carry different alleles.
B. Most alleles useful for DNA fingerprinting
differ on the basis of the number of repetitive
DNA sequences they contain.
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DNA Fingerprinting Basics
A DNA fingerprint is made by analyzing the sizes
of DNA fragments produced from a number of
different sites in the genome that vary in
length.
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The DNA Fragments Are Separated on the Basis of
Size
The technique is gel electrophoresis.
The pattern of DNA bands is compared between each
sample loaded on the gel.
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Gel Electophoresis

• A. Technique used to separate nucleic acids or
  proteins by size and charge.

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California Biology Content Standards

• California Content Standards
• 5 b. Students know how to apply base-pairing
  rules to explain precise copying of DNA.
• 5 c. Students know how genetic engineering
  (biotechnology) is used to produce novel
  biomedical and agricultural products.
• 5 e. Students know how exogenous DNA can be
  inserted into bacterial cells to alter their
  genetic makeup and support expression of new
  protein products.

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DNA Review

• A. Structure Double Helix
• B. Location Nucleus of Cell
• C. Function Blue print of Life Creation of
  Proteins/Amino Acids Transcription.
• D. Nitrogen Base ATCG
• E. Nucleotide Phosphate, Sugar (Deoxyribose),
  Nitrogen Base (ATCG).
• F. Base Pairing Rules A-T
• C-G

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Genetically Modified (GM) Food
Genetically Modified Cotton (contains a
bacterial gene for pest resistance)
Standard Cotton
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Genetically Modified (GM) Food
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What is Genetic Engineering?

• Genetic engineering is the technology for
  modifying the genetic information in a plant,
  animal or human in order to produce some desired
  trait or characteristic

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Genetic Engineering Vocabulary

• A. Restriction Enzymes molecular scissors are
  enzymes that cut DNA only at particular
  sequences.
• B. Plasmids are small circles of DNA found in
  bacteria.
• C. Plasmids are used to replicate a recombinant
  DNA.
• D. Vector - A vector is a small piece of DNA used
  to carry a gene of interest.

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Genetic Engineering Recombinant DNA Technology
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Restriction Enzymes are Enzymes that Cut DNA Only
at Particular Sequences
Different restriction enzymes have different
recognition sequences.
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DNA Cut by a Restriction EnzymesCan be Joined
Together in New Ways
These are recombinant DNAs and they often are
made of DNAs from different organisms.
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Plasmids are Used to Replicate a Recombinant DNA
A. Plasmids are small circles of DNA found in
bacteria.
B. Plasmids replicate independently of the
bacterial chromosome.
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Human Insulin Production by Bacteria

• 1. Isolate human cells and grow them in tissue
  culture.
• 2. Isolate DNA from the human cells.
• 3. Isolate plasmid DNA from a bacterium.
• 4. Use the same restriction enzyme to cut the
  plasmid DNA.
• 5. Mix the recombinant plasmid with bacteria.
• 6. Allow the new bacteria to incorporate the
  recombinant plasmid into the bacterial cell.
• 7. Grow trillions of new insulin producing
  bacteria (this is when cloning takes place).
• 8. A fermentor is used to grow recombinant
  bacteria.
• 9. Collect the bacteria, break open the cells
  and purify the insulin protein.

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Harnessing the Power of Recombinant DNA
Technology Human Insulin Production by Bacteria
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Human Insulin Production by Bacteria
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Human Insulin Production by Bacteria
Screening bacterial cells to learn which contain
the human insulin gene is the hard part.
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Route to the Production by Bacteria of Human
Insulin
This is the step when gene cloning takes place.
The single recombinant plasmid replicates within
a cell.
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Route to the Production by Bacteria of Human
Insulin
The final steps are to collect the bacteria,
break open the cells, and purify the insulin
protein expressed from the recombinant human
insulin gene.
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Overview of gene cloning.
Route to the Production by Bacteria of Human
Insulin.
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Genetic Engineering Insulin Production Overview
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Monday (March 12, 2012)Genetic Engineering

• Journal Question What is a plasmid?
• 1. Lecture II Genetic Engineering
• 2. Comprehensive Exam next Monday.

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Reviewing Genetic Engineering
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Banking Genes

• A. The massive Svalbard Global Seed Vault is
  built into the permafrost deep in a mountain on a
  remote arctic island in Norway

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Banking on Genes

• B. Built in 2008
• After receiving its first deposits, a
  doomsday seed vault on an Arctic island has
  amassed half a million seed samples, making it
  the worlds most diverse repository of crop
  seeds.

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Most Widely Used Genetically Modified Crops are

• 1. Cotton plants with a built-in resistance to
  insects.

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Most Widely Used Genetically Modified Crops are

• 2. Corn and Soybeans resistant to the herbicide
  Roundup.
• a. Allowing Farmers to employ no-till
  techniques to farming.

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Which country is the leader in plant
biotechnology?

• Answer China
• A. They have recently sequenced the rice genome.

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Problems with Genetic Engineering Technology

• 1. Environmental Problems
• 2. Food Safety
• 3. Access to the New Techniques

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Environmental Problems

• A. Pest resistant properties of transgenic
  crops.
• B. If pests have a broad exposure to the toxin
  or some other resistance incorporated into the
  plant, it is possible that they will develop
  resistance to the toxin and thus render it
  ineffective as an independent pesticide.

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Food Safety

• A. Food safety issues arise because transgenic
  crops contain proteins from different organisms
  and could trigger an unexpected allergic response
  to people who consume the food.

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Access to the New Techniques

• A. Relates to the developing world.
• B. Farmers in the developing countries are
  unable to afford the higher cost of the new
  genetically altered seeds.

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Other Types of Genetic Engineering

• 1. Transgenic Engineering
• a. Putting genetic information from one type of
  plant or animal into another.
• 2. Cloning
• a. Making exact genetic copies of an existing
  plant or animal.

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Transgenic Organisms

• A. An organism is called transgenic if it has
  genetic information added to it from a different
  type of organism.
• B. Viruses do something of this sort when they
  infect plants, animals or humans.
• C. Humans have begun to do this with plants and
  animals.

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Transgenic Organisms

• D. This is the work that is furthest along
• (1) Corn with its own insecticide.
• (2) Soybeans cotton resistant to
• herbicides.
• (3) Papayas resistant to viruses

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Transgenic Organisms

• F. Human genes have been inserted into
• (1) Bacteria (Prokaryotes)
• (2) Mice
• G. To produce various human proteins for
  treating diseases.

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Making Transgenic Mice
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Advantages of Transgenic Organisms

• A. Plants
• (1) More disease-resistant.
• (2) Larger yields.
• (3) More transportable.
• B. Animals
• (1) Make proteins for medicinal purposes.
• (2) Make organs for transplant to humans.

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Cloning Exact Copies

• A. A clone is an exact copy.
• B. In genetics, a clone is a genetic copy of
  another organism.
• C. Clones occur naturally
• Asexual breeding in plants lower animals
• Identical twins (triplets) in higher animals
• D. For centuries it has been known that simple
  animals worms starfish can be cloned by
  cutting them in half.
• E. This doesnt work for higher animals!
• F. Part of the problem is cell specialization
• Nerve, Bone, Muscle, etc.

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Cloning in the 20th Century

• A. We now realize that each specialized cell has
  all the genetic information, but much of it is
  turned off.
• B. Problem how to reset the program
• so this information is usable?
• C. Cloning of frogs successful in 1950s
• D. Cloning of livestock from fetal cells in
  1970s.

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Cloning in the 20th Century

• E. The human genome (an organisms genetic
  material) consists of 3 billion base pairs of DNA
  and about 30,000 genes.
• (1) 97 of our DNA does not code for protein
  product.
• -mostly consisting of repetitive sequences that
  never get transcribed.

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Cloning in the 20th Century Hello Dolly

• F. Clone from an adult sheep cell by Scots
  researchers under Ian Wilmut.
• G. Had only one success in 300 attempts.
• H. Dolly grew to maturity, and successfully
• had a lamb by natural means in 1998.
• I. But Dolly seems to be prematurely old.

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Cloning
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Cloning
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Genetic Engineering

• Genetic engineering (also known as genetic
  manipulation or GM) is not the same as cloning.
• -Though cloning techniques are used in genetic
  engineering, the two processes should not be
  confused.

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Genetic Engineering versus Cloning

• A. Cloning
• 1. Produces exact copies
• 2. Genes replicated within the
• same species.
• B. Genetic Engineering
• 1. Produces a totally unique
• set of genes.
• 2. Genes can be swapped across species.

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Selective Breeding versus Genetic Engineering

• A. In the past, humans have brought about change
  in the genetic make-up of organisms by means of
  selective breeding (artificial selection) i.e.
  Purebreds
• B. Genetic engineering brings about such change
  by scientifically altering an organism's genetic
  code.

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Genetic Engineering Overview

• 1. In genetic engineering enzymes are used to
  cut up and join together parts of the DNA of one
  organism, and insert them into the DNA of another
  organism.
• 2. In the resulting new organism the inserted
  genes will code for one or more new
  characteristics - for example producing a new
  substance, or performing a new function. The
  organism has been genetically re-engineered

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Other names for Genetic Engineering

• A. This technique is also known as gene splicing
  or recombinant DNA technology (because the DNA is
  recombined in the vector molecule.
• B. Vector - A vector is a small piece of DNA
  used to carry a gene of interest. Besides the
  gene being studied, a vector may contain elements
  which are used to help the gene integrate into a
  genome.

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Why does genetic engineering work?

• A. Genetic engineering works because there is
  only one code for life. The set of instructions
  for which a gene is responsible work whichever
  organism the gene is in, and whatever
  instructions that gene gives are carried out
  within the cells of the recipient.
• B. Theoretically the possibilities are limitless,
  although this sort of manipulation gives rise to
  strong feelings for and against.

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Applications of Genetic Engineering

• A. One field in which genetic engineering has
  had a huge impact is the mass production of
  insulin to help diabetics. Scientists have
  isolated the gene responsible for making human
  proteins, including the insulin hormone. This
  gene is inserted into the bacterial DNA, and the
  microbes then clone themselves rapidly, making
  identical copies of themselves, all with the new
  gene and all capable of making human insulin.
• B. This is a cheap way of producing sufficient
  quantities of exactly the right hormone, for
  everyone who needs it.

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Other Applications of Genetic Engineering

• C. Producing interferon, a human protein which
  stops viruses multiplying inside the body.
• D. Producing human growth hormone to treat growth
  abnormalities
• E. Blood clotting factor to treat hemophiliacs.
• F. Used in industry to produce enzymes for use
  in biological washing powder.
• G. Producing pest resistant crop varieties.
• H. Producing tomatoes and other produce that
  stay fresh much longer.

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Introduction to Genetic Engineering

• 1. With genetic engineering scientists directly
  manipulate genes.
• a. It frequently involves the use of recombinant
  DNA, which is composed of DNA segments from at
  least two different organisms.

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Commercial Applications

• 1. An example is the use of recombinant DNA
  technology to make interferon, a virus-destroying
  protein naturally produced by the human body.

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Production of Synthetic Interferon Involves

• 1. Isolating the human gene that codes for the
  interferon production.
• 2. Splicing this gene into a strand of bacterial
  DNA.
• 3. Inserting recombinant DNA into a bacterium.
• 4. Cloning the bacterium and collecting the
  product Interferon.

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Isolation of a Gene

• A. The first step in the process is isolating
  the human interferon gene.
• B. Genetic engineers use restriction enzymes,
  proteins that cut a DNA molecule into pieces.

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Isolation of a Gene

• C. The restriction enzyme EcoRI cuts DNA wherever
  the sequence C-T-T-A-A-G occurs.
• D. Other restriction enzymes cut DNA at different
  nucleotide sequences.
• E. By using the proper restriction enzymes
  scientists can cut the human interferon gene out
  of its chromosome.

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Gene Splicing
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Isolation of a Gene

• F. Once the gene for interferon is removed, it is
  separated from the rest of the DNA and then
  inserted into a strand of bacterial DNA.

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Gene Splicing

• A. Gene splicing is the process by which a gene
  from one organism is placed into the DNA of
  another organism.
• B. The human interferon gene is placed into the
  DNA of E. coli, the common bacterium of the human
  intestine.

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Gene Splicing

• C. In addition to a single, circular chromosome,
  E. coli contains a single, small ring of DNA
  called a plasmid.
• (Plasmid a single ring of DNA in bacteria)
• D. Human DNA is inserted into this plasmid.

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Gene Splicing

• E. The plasmid ring is removed from the bacterium
  and the opened with a restriction enzyme.
• F. The human interferon gene and the bacterial
  plasmid have sticky ends unpaired bases at
  each end of the DNA segment, where they were
  cleaved by restriction enzymes.

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Gene Splicing

• G. As the human DNA is spliced into the plasmid
  DNA, the unpaired bases of each bond readily.
• H. Consequently, a newly formed plasmid contains
  both human and bacterial DNA.

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Insertion, Cloning, and Collecting

• A. Once a DNA fragment is incorporated into a
  plasmid, the plasmid is inserted into another
  bacterium, which is then placed in a culture
  medium, where it divides and replicates.

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Insertion, Cloning, and Collecting

• B. Each time a bacterium divides a new copy of
  the plasmid DNA, which includes the human DNA
  gene, is created.
• C. This process by which the human gene is
  replicated is called gene cloning

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Insertion, Cloning, and Collecting

• D. Because E. coli can divide every 20 minutes,
  gene cloning is an efficient way to produce many
  copies of a specific genetic sequence.

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Insertion, Cloning, and Collecting

• E. The gene for human interferon is thus
  expressed in bacterial cultures and the resulting
  interferon protein is collected and eventually
  used by physicians.

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Genetically Modified (GM) Crops

• The ear of genetically engineered corn at top
  contains a toxin that kills worms.