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Genetic Engineering
- Archeological evidence shows that humans
  began cultivating plants and animals for food
  about 10,000 years ago

- Specific organisms were bred to improve
  future generations

- Simple form of applied genetics
CONTROLLED BREEDING


Manipulates the characteristics of the off
spring by selecting parents with specific
            phenotypic traits
Types of Controlled Breeding
1. Selective Breeding
     Process of choosing a few individuals to serve as parents
     for next generation

     - Luther Burbank: Californian plant
                       breeder (early 1900s)

     - responsible for 800 new plant varieties
                  Ex: seedless grape
                         Spineless cactus
                         Burbank potato
                         Daisies

     - Corn: bred for 7000 years
     - Small horses, dogs, cats: bred for different breeds
Types of Controlled Breeding
2. Inbreeding
   Process of crossing individuals with similar characteristics

   so those characteristics appear in offspring

  - purpose: to maintain or intensify desireable traits
  - individuals closely related (same species)
  - more risk of passing recessive genetic defects
          ex: poodles: joint deformities
              maine coon cats: heart murmurs
Types of Controlled Breeding
3. Hybridization
   Process of crossing individuals of different
   but related species
               - produces hybrids
               - hybrid vigor: offspring are hardier than parents
                               (grow faster, larger, and are healthier)
                     Ex: cattle - disease resistant
                         corn – yield 10 times more than old varieties




broccoflower          liger         zonkey        leopon       tangelo
Types of Controlled Breeding
1.   Induced Mutations (1927)

     Polyploidy: common in plants, deadly in animals
      - plants treated with chemicals to prevents cell plates
      - resulting cells have multiple sets of chromosomes
      - results in larger fruit and vegetables
• Techniques were forerunners modern
  genetic engineering

• Today geneticists engineer changes
  directly into an organismsDNA
GENETIC ENGINEERING
 (GENE SPLICING/GENE CLONING)

• Process of direct gene manipulation

• Goal: to introduce new characteristics into
        organisms to increase its usefulness

• Basic steps of genetic engineering involve:
      1. isolation of gene
      2. manipulation of gene/cloning gene -PCR
      3. reintroduction of DNA into model organisms

            animation: steps in cloning a gene
Genetic Engineering Techniques
I. Making Recombinant DNA

1. Restriction enzymes cut DNA into
   fragments that can be isolated and
   separated
   - very specific proteins, recognizes and
     cuts DNA at specific sequence into pieces
        Ex: EcoRI – cuts DNA whenever
            C-T-T-A-A-G sequence occurs

2. Production of recombinant DNA
   - DNA composed of fragments of DNA
     segments from at least two different
     organisms
   - restriction enzymes cut bacterial
     plasmids (extra circular DNA molecules
     in bacteria)
   - plasmids have “sticky ends”
     (unpaired bases)
   - original DNA is attached to plasmid
     sticky ends
Genetic Engineering Techniques
3. reintroduction of DNA into bacterial
   vector
  - recombinant DNA taken up with bacterial
    DNA and now produced by bacterial cell
  - recombinant DNA is isolated and CLONED
  - PCR (polymerase chain reaction) induced
  - purpose: to make 1000s of recombinant
             plasmids
                 PCR animation

• DNA sequencing
  - Process of reading exact order of bases in
    fragment of DNA
  - makes it possible for scientists to make
    sure gene of interest has been cloned
              rDNA: animation
Genetic Engineering Techniques
II. Microinjection
   - Process of injecting genetic
     material containing the new
     gene into the recipient cell
        - direct gene transfer, no
          use of vectors
        - in large cells done with
          fine tipped glass needle
        - somehow injected genes
          find the host cell genes
          and incorporate
          themselves among them

          Ex: most common
              method of making
              genetically altered
              mice
Genetic Engineering Techniques
I.     Bioballistics                   I.     Electro and Chemical
                                                  Poration
     - projectile methods that use
        metal slivers to deliver the        - process that creates pores
        genetic material to the                or holes in the cell
        interior of the cell.                  membrane to allow direct
                                               entry of new genes
     - small slivers coated with
       genetic material                     - done by bathing cells in
                                               solutions of special
                                               chemicals or weak electric
     - once in the cell, genetic
                                               current
       material is transported to
       nucleus where it is
       incorporated among the host
       genes
TRANSGENIC ORGANISMS /
GENETICALLY MODIFIED ORGANISMS
            (GMO’s)

• organisms that contain foreign genes

Transgenesis
  The use of recombinant DNA techniques to
  introduce new characters (ie. genes) into
  organisms (including humans) that were not
  present previously.
Types GMO’s
I. Bacteria

-human DNA inserted in
 bacterial plasmid

-recombinant produces large
 volumes of proteins

  Ex: human growth hormone
      (HGH), insulin
Types GMO’s
II. Plants
 - 1986: Howell
     - inserted gene to producer enzyme
       luciferase (fireflies)into tobacco plant
           - enzyme in fireflies
           - only produces enzyme in
           conditions without light
            (gene expression)

    - cloned cells reproduced tobacco plant that
      glowed in the dark
Making Transgenic Crops
                        Steps
               2. extracting DNA
               3. cloning a gene of
                  interest
               4. designing the gene
                  for plant infiltration
               5. transformation
               6. plant breeding
Other Transgenic Plants
BT corn
(bacillus thuringiensis)

Soil bacterium that resists
insecticidal toxins: makes
corn resistant to pests and
less need for pesticides
Other Transgenic Plants

             Calgene tomato
             “Flavr Savr”

             stays fresh longer
             because enzyme to break
             down pectin are reduced
             by genetic alteration
Types GMO’s
III. Animals
Transgenic sheep:

human gene for milk
production inserted into
sheep and now produces
proteins of human milk
Oncomice
(cancer mice)

Used in medical
research to find cures
for cancer
13 genetic engineering bw
Other examples of transgenic animals:

Transgenic chickens:
- grow faster and larger in close quarters
- produce more protein in their egg whites
- produce human protein drugs in their eggs

             Transgenic pigs:
             - produced by fertilizing normal eggs with
               sperm cells that have incorporated foreign
               DNA

      - may someday be able to produce transgenic pigs
        that can serve as a source of transplanted organs
        for humans
                         video
Cloning Animals
1997 Wilmut (Scottish)
                         - cloned sheep “Dolly”

                         - process: nucleus of egg is replaced
                           with nucleus of adult , resultant
                           organism identical to adult

                         - now companies are making cloning
                           available to pet owners for their
                           beloved pets who die

                         - is this ethical?



                                                  Genetic Savings and Clone
Ethics of GMOs and Cloning
• Imagine cloning humans to make armies.

• Could clones and GMOs be patented?

• Are genetically modified livestock and agriculture
  harmful?

• Would only perfect genetically engineered infants be
  desirable.
APPLICATIONS OF G.E.
The first genetic fingerprint      1. DNA Fingerprinting
           1984                       (used in forensics)

                                    Process of identifying and
                                    distinguishing DNA of individuals
                                    - each person has unique repeat
                                     sequences and numbers of
                                      non coding introns
                                    - sample is taken and DNA is
                                      extracted
                                    - fragments of repeats of introns
                                      are labeled and put into a
                                      gel electrophoresis where they
                                      separate
                                    - create banding patterns which
Electrophoresis Animation Detail      are unique for every human
APPLICATIONS OF G.E.
- Once banding patterns are apparent they are transferred to a
  Southern Blot for identification




                       Southern blot animation
Applications of Genetic Engineering
       DNA Fingerprinting
Applications of Genetic Engineering
       DNA Fingerprinting




    Restriction fragment length polymorphisms
Applications of Genetic Engineering
       DNA Fingerprinting
APPLICATIONS OF G.E.
     Microarray
APPLICATIONS OF G.E.
•   Gene therapy
    Creating transgenic
    humans to erase
    genetic diseases

    - recombinant DNA
    put directly into human
    cells or through virus
    vectors
APPLICATIONS OF G.E.
                                                                1. Pollution control

                                                                  - genetically altered
                                                                  bacteria used to
                                                                  decompose garbage
                                                                  sewage, and petroleum
                                                                  products

Unlike the left tower, which uses chemicals, the tower on the
right at this wastewater-treatment plant now uses bacteria-
covered foam blocks (inset) to eliminate the hydrogen sulfide
bubbling from treated sewage.
Deshusses/PNAS
APPLICATIONS OF G.E.
1.   Medicines and Vaccines produced by
     bacteria and viruses

     - E coli: used to make human insulin

     - hamster cell cultures: used to
       make TPA (tissue plasminogen
       activator)- dissolves blood clots in
       heart attacks

     - EPO (erythropoiten): increases
       red blood cell production

     - interferon: fights viral infections,
       increases immunity

     - vaccines

                   vaccine animation
APPLICATIONS OF G.E.

5. Agriculture and livestock

 - hardier, disease and environmentally
   resistant fruits and vegetables

 - larger sturdier animals
HUMAN GENOME PROJECT




• begun in 1990: coordinated by US Dept of Energy and NIH
• purpose: - to identify the 20-25,000 genes in human DNA
            - to determine sequences of 3 billion DNA base pairs
            - to license info to biotech companies to foster new medical
              applications
• international: US, England, China, France, Germany, Japan
• used combined genomes of anonymous small number of people
Study for the test !

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13 genetic engineering bw

  • 2. - Archeological evidence shows that humans began cultivating plants and animals for food about 10,000 years ago - Specific organisms were bred to improve future generations - Simple form of applied genetics
  • 3. CONTROLLED BREEDING Manipulates the characteristics of the off spring by selecting parents with specific phenotypic traits
  • 4. Types of Controlled Breeding 1. Selective Breeding Process of choosing a few individuals to serve as parents for next generation - Luther Burbank: Californian plant breeder (early 1900s) - responsible for 800 new plant varieties Ex: seedless grape Spineless cactus Burbank potato Daisies - Corn: bred for 7000 years - Small horses, dogs, cats: bred for different breeds
  • 5. Types of Controlled Breeding 2. Inbreeding Process of crossing individuals with similar characteristics so those characteristics appear in offspring - purpose: to maintain or intensify desireable traits - individuals closely related (same species) - more risk of passing recessive genetic defects ex: poodles: joint deformities maine coon cats: heart murmurs
  • 6. Types of Controlled Breeding 3. Hybridization Process of crossing individuals of different but related species - produces hybrids - hybrid vigor: offspring are hardier than parents (grow faster, larger, and are healthier) Ex: cattle - disease resistant corn – yield 10 times more than old varieties broccoflower liger zonkey leopon tangelo
  • 7. Types of Controlled Breeding 1. Induced Mutations (1927) Polyploidy: common in plants, deadly in animals - plants treated with chemicals to prevents cell plates - resulting cells have multiple sets of chromosomes - results in larger fruit and vegetables
  • 8. • Techniques were forerunners modern genetic engineering • Today geneticists engineer changes directly into an organismsDNA
  • 9. GENETIC ENGINEERING (GENE SPLICING/GENE CLONING) • Process of direct gene manipulation • Goal: to introduce new characteristics into organisms to increase its usefulness • Basic steps of genetic engineering involve: 1. isolation of gene 2. manipulation of gene/cloning gene -PCR 3. reintroduction of DNA into model organisms animation: steps in cloning a gene
  • 10. Genetic Engineering Techniques I. Making Recombinant DNA 1. Restriction enzymes cut DNA into fragments that can be isolated and separated - very specific proteins, recognizes and cuts DNA at specific sequence into pieces Ex: EcoRI – cuts DNA whenever C-T-T-A-A-G sequence occurs 2. Production of recombinant DNA - DNA composed of fragments of DNA segments from at least two different organisms - restriction enzymes cut bacterial plasmids (extra circular DNA molecules in bacteria) - plasmids have “sticky ends” (unpaired bases) - original DNA is attached to plasmid sticky ends
  • 11. Genetic Engineering Techniques 3. reintroduction of DNA into bacterial vector - recombinant DNA taken up with bacterial DNA and now produced by bacterial cell - recombinant DNA is isolated and CLONED - PCR (polymerase chain reaction) induced - purpose: to make 1000s of recombinant plasmids PCR animation • DNA sequencing - Process of reading exact order of bases in fragment of DNA - makes it possible for scientists to make sure gene of interest has been cloned rDNA: animation
  • 12. Genetic Engineering Techniques II. Microinjection - Process of injecting genetic material containing the new gene into the recipient cell - direct gene transfer, no use of vectors - in large cells done with fine tipped glass needle - somehow injected genes find the host cell genes and incorporate themselves among them Ex: most common method of making genetically altered mice
  • 13. Genetic Engineering Techniques I. Bioballistics I. Electro and Chemical Poration - projectile methods that use metal slivers to deliver the - process that creates pores genetic material to the or holes in the cell interior of the cell. membrane to allow direct entry of new genes - small slivers coated with genetic material - done by bathing cells in solutions of special chemicals or weak electric - once in the cell, genetic current material is transported to nucleus where it is incorporated among the host genes
  • 14. TRANSGENIC ORGANISMS / GENETICALLY MODIFIED ORGANISMS (GMO’s) • organisms that contain foreign genes Transgenesis The use of recombinant DNA techniques to introduce new characters (ie. genes) into organisms (including humans) that were not present previously.
  • 15. Types GMO’s I. Bacteria -human DNA inserted in bacterial plasmid -recombinant produces large volumes of proteins Ex: human growth hormone (HGH), insulin
  • 16. Types GMO’s II. Plants - 1986: Howell - inserted gene to producer enzyme luciferase (fireflies)into tobacco plant - enzyme in fireflies - only produces enzyme in conditions without light (gene expression) - cloned cells reproduced tobacco plant that glowed in the dark
  • 17. Making Transgenic Crops Steps 2. extracting DNA 3. cloning a gene of interest 4. designing the gene for plant infiltration 5. transformation 6. plant breeding
  • 18. Other Transgenic Plants BT corn (bacillus thuringiensis) Soil bacterium that resists insecticidal toxins: makes corn resistant to pests and less need for pesticides
  • 19. Other Transgenic Plants Calgene tomato “Flavr Savr” stays fresh longer because enzyme to break down pectin are reduced by genetic alteration
  • 21. Transgenic sheep: human gene for milk production inserted into sheep and now produces proteins of human milk
  • 22. Oncomice (cancer mice) Used in medical research to find cures for cancer
  • 24. Other examples of transgenic animals: Transgenic chickens: - grow faster and larger in close quarters - produce more protein in their egg whites - produce human protein drugs in their eggs Transgenic pigs: - produced by fertilizing normal eggs with sperm cells that have incorporated foreign DNA - may someday be able to produce transgenic pigs that can serve as a source of transplanted organs for humans video
  • 25. Cloning Animals 1997 Wilmut (Scottish) - cloned sheep “Dolly” - process: nucleus of egg is replaced with nucleus of adult , resultant organism identical to adult - now companies are making cloning available to pet owners for their beloved pets who die - is this ethical? Genetic Savings and Clone
  • 26. Ethics of GMOs and Cloning • Imagine cloning humans to make armies. • Could clones and GMOs be patented? • Are genetically modified livestock and agriculture harmful? • Would only perfect genetically engineered infants be desirable.
  • 27. APPLICATIONS OF G.E. The first genetic fingerprint 1. DNA Fingerprinting 1984 (used in forensics) Process of identifying and distinguishing DNA of individuals - each person has unique repeat sequences and numbers of non coding introns - sample is taken and DNA is extracted - fragments of repeats of introns are labeled and put into a gel electrophoresis where they separate - create banding patterns which Electrophoresis Animation Detail are unique for every human
  • 28. APPLICATIONS OF G.E. - Once banding patterns are apparent they are transferred to a Southern Blot for identification Southern blot animation
  • 29. Applications of Genetic Engineering DNA Fingerprinting
  • 30. Applications of Genetic Engineering DNA Fingerprinting Restriction fragment length polymorphisms
  • 31. Applications of Genetic Engineering DNA Fingerprinting
  • 32. APPLICATIONS OF G.E. Microarray
  • 33. APPLICATIONS OF G.E. • Gene therapy Creating transgenic humans to erase genetic diseases - recombinant DNA put directly into human cells or through virus vectors
  • 34. APPLICATIONS OF G.E. 1. Pollution control - genetically altered bacteria used to decompose garbage sewage, and petroleum products Unlike the left tower, which uses chemicals, the tower on the right at this wastewater-treatment plant now uses bacteria- covered foam blocks (inset) to eliminate the hydrogen sulfide bubbling from treated sewage. Deshusses/PNAS
  • 35. APPLICATIONS OF G.E. 1. Medicines and Vaccines produced by bacteria and viruses - E coli: used to make human insulin - hamster cell cultures: used to make TPA (tissue plasminogen activator)- dissolves blood clots in heart attacks - EPO (erythropoiten): increases red blood cell production - interferon: fights viral infections, increases immunity - vaccines vaccine animation
  • 36. APPLICATIONS OF G.E. 5. Agriculture and livestock - hardier, disease and environmentally resistant fruits and vegetables - larger sturdier animals
  • 37. HUMAN GENOME PROJECT • begun in 1990: coordinated by US Dept of Energy and NIH • purpose: - to identify the 20-25,000 genes in human DNA - to determine sequences of 3 billion DNA base pairs - to license info to biotech companies to foster new medical applications • international: US, England, China, France, Germany, Japan • used combined genomes of anonymous small number of people
  • 38. Study for the test !