Biotechnology

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Biotechnology

• bios life technos tool logos study of
• Biotechnology The Study of Living Tools

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1. Timeline

• 4000 BC Egyptians use yeasts for bread and wine
• 1750 BC Sumerians brew beer
• 1500 AD Aztecs make cakes from Spirulina algae
• 1917 Biotechnology term coined
• 1972 Hamilton Smith discovers first restriction
  enzyme
• 1973 Stanley Cohen made first transgenic organism
  with gene from African clawed toad into
  bacteria
• 1978 Louise Brown, first test tube baby born
• 1981 PCR Invented by Kary Mullis
• Genentech releases (Humulin?) human insulin
• 1984 PCR used by Alec Jeffries in DNA
  fingerprinting
• EPA approves release of genetically engineered
  tobacco
• 1990 Pfizer introduces Chymosin (Rennin)
• Michael Crichtons Jurassic Park published
• 1993 FDA approval of Monsantos rBGH/rBST
• 1994 Calgene introduces Flavr-Savr? Tomato
• 1995 O.J. Simpson Trial
• 1996 Sequence completed for S. cervisiae
• 1997 Cloning of Dolly at Roslin Institute
• 1998 First animal genome sequenced C. elegans

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Selective Breeding

• a. Luther Burbank
• disease resistant Burbank potato
• fight blight, etc in Ireland
• b. Norman Borlaug of International Maize
• Wheat Research Center in Mexico
• (received Nobel Prize)
• Crossed short-stemmed wheat with
• Mexicos best wheat
• Govt of India requested seeds,
• as tall wheat plants falling over
• Increased wheat production from 12 million metric
  tons in 1965, to 20 million in 1970, 37 million
  in 1982
• c. Hybridization
• Dissimilar individuals mate to hope to get
  desired traits
• Burbank Popular Shasta Daisies
• d. Inbreeding
• Keep desired traits
• Brings recessive traits too (joints in golden
  retrievers)

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Increasing Variation

• a. Inducing mutations in bacteria
• Radiation, chemicals, r-strategists
• Can clean up oil spills (bioremediation)
• b. Polyploidy (extra chromosomes)
• Usually fatal in animals
• Bigger, stronger, sexier plants (bananas, citrus
  fruits, day lilies)

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DNA Manipulation

• 1. Cutting Separating
• a. Restriction Enzymes
• Proteins from bacteria that cut DNA at specific
  points
• Cut at palindromes
• Evolved as viral defenses
• Can have blunt or sticky ends
• Can be spliced into DNA cut with same RE

Naming of EcoR1 E from genus of organism where
found (Escheria) co first 2 letters of species
name (coli) R Strain (RY13) 1 Order discovered
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Restriction Enzymes
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(Cutting Separating, Contd)

• b. Gel Electrophoresis
• Migration of charged particles under electric
  field
• DNA has negatively charged phosphate ends
• (-) electrode repels DNA, () electrode attracts
  DNA
• 1 agarose gel (natural colloid from seaweed)
• agarose is convoluted like a sieve
• TBE solution is electrolytic solution
• Migration of DNA molecules move through gel at
  different rates (bigger slower, smaller
  faster)
• DNA is stained to see bands (Ethidium Bromide)
• Usually a marker is used (of known fragment sizes)

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Equipment
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Digest Separation
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(Cutting Separating, Contd)

• c. Restriction Mapping
• Use of various restriction enzymes
• Gel digests show size of fragments
• Patterns of digests can create restriction map
• pUK 1 plasmid

Gel Digest Restriction Map
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• Restriction digest of plasmid DNA from
  Escherichia coli
• run on a 1 agarose gel and stained with ethidium
  bromide
• Lane 1 (far left) is a kilobase DNA ladder
• Lane 2 is the uncut plasmid DNA
• Lane 3 is a single digestion of the plasmid with
  the EcoRI
• Lane 4 is also a single digestion, but with XhoI
• Lane 5 (far right) is a double digestion - both
  EcoRI and XhoI

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DNA Manipulation, Contd

• 2. Identifying Genes - Southern Blot (named after
  Ed Southern)
• a. DNA cut with REs
• b. Separated by Gel Electrophoresis
• c. DNA blotted to filter paper and probe is
  added
• d. Only DNA fragments with identified gene bind
  to probe

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Southern Blot
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DNA Manipulation, Contd

• 3. Nucleotide Sequencing (fluorescent
  dye-terminator cycle sequencing)
• a. Unknown Single Stranded DNA put in test tube
• b. DNA polymerase and nucleotide bases (dNTPs)
  added
• c. Small number of bases with flourescent dye
  attached (ddNTPs)
• d. Each time dye-labeled nucleotide binds to
  fragment, synthesis stops
• e. Ultimately yields DNA strands of different
  lengths
• f. Separated (by gel electrophoresis)
• g. Color of bands tells sequence (read by laser
  detector, computer software analyzes)

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Laser Readout
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ABI Prism
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DNA Manipulation, Contd

• 4. Making Copies (Polymerase Chain Reaction or
  PCR)
• a. Small amount of double stranded DNA
• b. Heated to separate, then cooled
• c. Add DNA polymerase, primers (short pieces of
  artificial DNA), and free nucleotides
• Taq polymerase (from Thermophilus aquaticus)
• Isolated from hot springs in Yellowstone
• Can withstand hot temperatures
• d. DNA poly attaches nucleotides to primers
• e. REPEATED many times (5 minute cycles)
• f. Use of a thermal cycler

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DNA Manipulation, Contd

• 5. Recombinant DNA
• a. Use REs to cut out gene
• b. Cut host DNA with same RE
• c. Need a vector to incorporate into host
• d. Screen for effective transfer
• e. If successful, then results in Transgenic
  Organism
• f. First done by Steven Howell by inserting
  luciferase gene (for glowing) from firefly into
  tobacco plant
• g. Transgenic Organisms
• 1. Bacteria
• Transformation with plasmid
• Plasmid ring of DNA used to transfer genes

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• 2. Plants
• Bacterium
• Has small DNA plasmid causes tumors
• Inactivation of tumor gene
• Insertion of foreign DNA
• May uptake DNA if cell walls removed
• Gene gun!
• 3. Animals
• Viral vectors

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• 4. Nuclear Transfer
• Can inject DNA into large egg nuclei
• Enzymes used to repair and recombine DNA in cell
  help insert foreign DNA

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• 5. Scientists may also insert marker gene to tell
  if procedure worked
• ampicillin resistance (bacteria then grown on
  nutrient medium with ampicillin)
• glowing gene from jellyfish (produces GFP)

To determine what turns on color in wings,
University of Buffalo (NY) biologists inserted a
marker gene from jellyfish into African
butterflies resulting in fluorescent green eyes
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Using Glowing Markers
Fruit Fly Embryo
Glowing Tobacco!!
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• 6. Knockout Mice
• Scientists transfer a defective version of a gene
  they want to study into stem cells
• The defective gene knocks out the normal gene,
  and scientists can examine the effects of the
  disabled gene on the resulting young mouse.
• Using gene targeting, researchers can transfer
  human disease genes into embryonic stem cells to
  make mouse models of many human ailments

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• 7. Whole Genome Analysis
• Allows analysis of multiple genes in various
  conditions
• Complexity does not come from the number of
  genes, but from the way in which they are used
  (Gerald Rubin, HHMI VP)
• Gene Chips (Affymetrix)
• ½ square inch glass with short DNA fragments
• 200,000 each
• Microarrayer
• Robot designed by Patrick Brown _at_ Stanford
• Can analyze 6,000 genes in yeast simultaneously
• Make your own for 25,000

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A Microarrayer Shows How Genes in a Yeast Cell
Respond to Different Types of Stress
http//www.hhmi.org/genesweshare/a110.html
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US GOV vs. Celera Genomics
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Uses

• 1. Human Genome Project
• a. Attempt to map all of human genome!
• b. Begun in 1999, working draft Feb. 2001,
  finished 2003 (3 years ahead of schedule!)
• c. Collaboration of 20 labs in 6 countries
• d. Competition with Craig Ventnor, Celera
  Genomics
• e. Discoveries
• 3.2 billion base pairs
• only 30-40,000 genes
• over 120,000 unique mRNA molecules
• only 1-1.5 of human DNA codes for proteins
• Each cell has 6 ft of DNA 1 inch of exons to be
  transcribed
• Most of genome is Junk DNA
• Genes not evenly distributed
• Chromosome 19 packed with genes
• Large chromosomes 4 8 have few transcribed genes

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Types of Genetic Maps
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pGLO Plasmid Map
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• pGLO Sequence
• ATCGATGCATAATGTGCCTGTCAAATGGACGAAGCAGGGATTCTGCAAA
  CCCTATGCTACTCCGTCAAGCCGTCAATTGTCTGATTCGTTACCAATTAT
  GACAACTTGACGGCTACATCATTCACTTTTTCTTCACAACCGGCACGGAA
  CTCGCTCGGGCTGGCCCCGGTGCATTTTTTAAATACCCGCGAGAAATAGA
  GTTGATCGTCAAAACCAACATTGCGACCGACGGTGGCGATAGGCATCCGG
  GTGGTGCTCAAAAGCAGCTTCGCCTGGCTGATACGTTGGTCCTCGCGCCA
  GCTTAAGACGCTAATCCCTAACTGCTGGCGGAAAAGATGTGACAGACGCG
  ACGGCGACAAGCAAACATGCTGTGCGACGCTGGCGATATCAAAATTGCTG
  TCTGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTACCCGATTATC
  CATCGGTGGATGGAGCGACTCGTTAATCGCTTCCATGCGCCGCAGTAACA
  ATTGCTCAAGCAGATTTATCGCCAGCAGCTCCGAATAGCGCCCTTCCCCT
  TGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAATGCGGCTGGTG
  CGCTTCATCCGGGCGAAAGAACCCCGTATTGGCAAATATTGACGGCCAGT
  TAAGCCATTCATGCCAGTAGGCGCGCGGACGAAAGTAAACCCACTGGTGA
  TACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAATCTCTCCTGG
  CGGGAACAGCAAAATATCACCCGGTCGGCAAACAAATTCTCGTCCCTGAT
  TTTTCACCACCCCCTGACCGCGAATGGTGAGATTGAGAATATAACCTTTC
  ATTCCCAGCGGTCGGTCGATAAAAAAATCGAGATAACCGTTGGCCTCAAT
  CGGCGTTAAACCCGCCACCAGATGGGCATTAAACGAGTATCCCGGCAGCA
  GGGGATCATTTTGCGCTTCAGCCATACTTTTCATACTCCCGCCATTCAGA
  GAAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTGCGTCTT
  TTACTGGCTCTTCTCGCTAACCAAACCGGTAACCCCGCTTATTAAAAGCA
  TTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCGTAACAAAAGT
  GTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGCACGGCGTCAC
  ACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCGGATCCTACCT
  GACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCCGTTTTTTTGG
  GCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGCTAG
  CAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAG
  ATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGT
  GATGCTACATACGGAAAGCTTACCCTTAAATTTATTTGCACTACTGGAAA
  ACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGGTGTTC
  AATGCTTTTCCCGTTATCCGGATCATATGAAACGGCATGACTTTTTCAAG
  AGTGCCATGCCCGAAGGTTATGTACAGGAACGCACTATATCTTTCAAAGA
  TGACGGGAACTACAAGACGCGTGCTGAAGTCAAGTTTGAAGGTGATACCC
  TTGTTAATCGTATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAAC
  ATTCTCGGACACAAACTCGAGTACAACTATAACTCACACAATGTATACAT
  CACGGCAGACAAACAAAAGAATGGAATCAAAGCTAACTTCAAAATTCGCC
  ACAACATTGAAGATGGATCCGTTCAACTAGCAGACCATTATCAACAAAAT
  ACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTC
  GACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGCGTGACCACATGG
  TCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACACATGGCATGGATGAG
  CTCTACAAATAATGAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCG
  ACCTGCAGGCATGCAAGCTTGGCTGTTTTGGCGGATGAGAGAAGATTTTC
  AGCCTGATACAGATTAAATCAGAACGCAGAAGCGGTCTGATAAAACAGAA
  TTTGCCTGGCGGCAGTAGCGCGGTGGTCCCACCTGACCCCATGCCGAACT
  CAGAAGTGAAACGCCGTAGCGCCGATGGTAGTGTGGGGTCCCCCATGCGA
  GAGTAGGGAACTGCCAGGCATCAAATAAAACGAAAGGCTCAGTGCAAAGA
  CTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTA
  GGACAAATCCGCCGGGAGCGGATTTGAACGTTGCGAAGCAACGGCCCGGA
  GGGTGGCGGGCAGGACGCCCGCCATAAACTGCCAGGCATCAAATTAAGCA
  GAAGGCCATCCTGACGGATGGCCTTTTTGCGTTTCTACAAACTCTTTGTT
  TATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCC
  TGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACA
  TTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTT
  TTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTG
  GGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCT
  TGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAG
  TTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAA
  CTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC
  AGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCA
  GTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACA
  ACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGA
  TCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATAC
  CAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTG
  CGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATT
  AATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGG
  CCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGT
  GGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCG
  TATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAA
  ATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTG
  TCAGACCAAGTTTACTCATATATACTTTAGATTGATTTACGCGCCCTGTA
  GCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCT
  ACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTT
  TCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCC
  CTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTT
  GATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTT
  TCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCC
  AAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAA
  GGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACA
  AAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTAAAAGG
  ATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACG
  TGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGAT
  CTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAA
  AAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAAC
  TCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTG
  TCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCA
  CCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAG
  TGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGG
  ATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGC
  TTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTG
  AGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAA
  GCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC
  GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCG
  TCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCA
  GCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCAC
  ATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGC
  CTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCG
  AGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTT
  ACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAAT
  CTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACG
  TGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGC
  CCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACC
  GTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACG
  CGCGAGGCAGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCT
  GCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGC
  CCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCAC
  CTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCTAA
  TTCTCATGTTTGACAGCTTATC

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• 2. DNA Fingerprinting
• a. History
• Lynda Mann in England
• Raped and Murdered, Nov. 22, 1983
• Local dishwasher questioned pleads guilty to
  similar case
• Alec Jeffries uses new method of PCR for
  identification, exonerates dishwasher of both
  crimes
• Every man in area fingerprinted by DNA, no
  matches
• Colin Pitchfork finally caught and tested
  positive (friend went in to fake test for him)
• Plant Witness
• Murder case in Phoenix, Arizona
• Pager found at crime scene led police to suspect
  (said that victim had robbed him)
• Palo Verde pods in truck yielded DNA that matched
  with trees at crime scene

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• b. Uses
• Identification and exoneration of rapists,
  criminals
• Paternity cases (Jefferson/Sally Hemmings)
• Identification of body parts
• Supposed heart of Louis XVIII (child king who
  died in prison) compared to hair from Marie
  Antoinette
• ID of bodies in mass graves in Guatemala (from
  civil war)
• Genetic testing (blood stain on Lincolns jacket
  tested for Marfans syndrome)
• Migration patterns
• ID tags for children, pets
• ID of endangered/protected species
• Food authentication, such as in wine and caviar
• Authentication of official 2000 Summer Olympic
  goods (sections of DNA taken from several unnamed
  Australian athletes added to ink used to mark all
  items)

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National Geographic March 2006
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• c. Use of RFLPs (restriction fragment length
  polymorphisms)
• Procedure
• Restriction enzymes cut DNA differently
• Fragments separated with GE
• Probed and exposed to X-ray film
• Screening for Sickle Cell Anemia
• Point mutation of CAG (betaA gene) to CTG (betaS
  gene) SNP or single nucleotide polymorphism
• Produces valine instead of glutamic acid in
  hemoglobin molecule
• REs cut DNA differently
• Probe attaches to specific sequence
• Affected large fragment
• Pedigree shows family son with sickle-cell
  anemia
• Electrophoresis pattern below each child

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RFLP
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• d. DNA Typing
• Small percentage of DNA different from person to
  person (less than 1/10th of 1)
• Variable regions used for comparison

Gel Lanes MARKERS VICTIM EVIDENCE 1 (semen stain
left on the victim's clothing) EVIDENCE 2
(semen from the vagina of the rape victim)
SUSPECT 1 SUSPECT 2 CONTROL (check to see if
probes are working)

• Results
• Suspect 2 can be clearly ruled out
• Suspect 1 MAY be guilty (probability that 6
  alleles match is 1 in 4056)
• Suspects NOT picked at random Evidence used in
  conjunction with
• More probes (alleles) the better 14 chance of
  match is 1 in 268 million

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• 3. Transgenic Organisms
• a. Medicine
• 1. Isolate gene for protein needed for medicine
• 2. Inserted into bacteria, will make protein in
  normal course of its life cultured in bioreactor

Humulin? (Human Insulin)
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Genetically Engineered Medicines

• Erythropoetin Anemia
• Growth Factors Burns, Ulcers
• Human Growth
• Hormone (HGH) Dwarfism
• Insulin Diabetes
• Interferons Viral Infections, Cancer
• Taxol Ovarian Cancer

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• 3. Inserted into animals (livestock)
• Dolly cloned in 1997 by Ian Wilmut in Scotland
• Potential to produce protein in milk
• Cloning done once, then livestock reproduce
  regularly to pass on gene FARMacy

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• b. Agriculture
• 1. Improving crops
• Crops resistant to biodegradable weedkiller
  (glyphosate)
• Kills only weeds
• Half of 72 million acres of Soybeans in 2000
• Flavr-Savr? tomato last longer
• Genetically enhanced rice with more Vitamin A

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• Concerns
• Natural resistance to enhancement
• Jumping genes
• Allergies
• 2. Improving livestock growth hormone added to
  cow (bGH/bST)
• c. Bioremediation - microorganisms engineered to
  feed on toxic or hazardous materials
• 1. Pseudomonas bacteria engineered to degrade
  polyhalogentated compounds (pollutants)
• 2. E. coli engineered to clean up mercury

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Bioremediation Cartoon
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4. RNA interference (RNAi)

• post-transcriptional gene silencing
• double stranded RNA (dsRNA) causes
  sequence-specific degradation of homogolous
  mRNAsequences
• First found in C. elegans

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The Future?

• Alternatives to antibiotics to protect livestock
  against bacterial infections
• 2nd Gen of GM crops is expected that could
  eliminate of allergens in food, increase
  nutritional content, and lower fat and oil levels
• 3rd Gen GM crops may have properties like salt
  tolerance, drought resistance, drugs and vaccines
  within them, and plastic starter chemicals to
  create bioplastics
• http//www.biotechnologyonline.gov.au/foodag/timel
  ine.cfm