1. Biotechnology
Use of Natural Biological Systems to Produce a Product or Provide a Desired Process

2. Recombinant DNA Technology
A set of techniques used to produce large quantities of a gene and its product
Recombinant DNA = DNA produced by joining segments of DNA from different sources
eg. To produce human insulin, scientists have combined bacterial DNA + human DNA

3. Tools for Producing Recombinant DNA
Restriction enzymes: enzymes that cleave the DNA double helix at specific nucleotide sequences

4. Use of the Restriction Enzyme Bam H1
5’— G G A T C C — 3’
3’— C C T A G G — 5’
Results in
sticky end
5’— G G A T C C — 3’
3’— C C T A G G — 5’

5. Tools for Producing Recombinant DNA
Vector: carrier of DNA; can be virus or plasmid
Plasmid: extrachromosomal, independently replicating, small circular DNA molecule

6. Producing Recombinant DNA
restriction enzyme
Mix together
Add DNA Ligase
Many recombinant DNA
molecules are produced,
each with a different piece of source DNA
Treat source DNA with restriction enzyme
Treat plasmid DNA with same enzyme
restriction enzyme
Transform bacterial cells
Each bacterial cell carries a different recombinant plasmid

7. Tools for Producing Recombinant DNA
Probe: sequence of DNA that is complementary to the gene of interest; Used to locate a copy of the gene by hybridization
AGCTTAGCGAT
TCGAATCGCTA
Denature DNA by heating
AGCTTAGCGAT
TCGAATCGCTA
AATCGC
Add Probe
Probe Binds to gene

8. Using the Probe to Find the Gene of Interest

9. Biotechnological Methods: PCR http://www. dnalc
PCR = Polymerase Chain Reaction
Amplifies a specific region in the DNA
Used for identification, especially if the amount of DNA is small
Uses repeated cycles of heating to denature DNA and cooling to synthesize new DNA
Involves the use of
---Taq polymerase (a DNA polymerase that withstands heat)
---primers to begin synthesis

10. DNA Fingerprinting with PCR

11. DNA Fingerprinting for Paternity Testing

12. DNA Fingerprinting in Forensics1 2 3
C S R C I E
M N E E 4 5 6 7
Suspects
Suspects

13. DNA sequence for gene from cress plant
Biotechnological Methods DNA Sequencing: Determining the Order of Nucleotides
Arrows show differences in DNA sequences related to inherited breast cancer
DNA sequence for gene from cress plant

14. Human Genome Project
An effort to determine the sequences of all human genes
Genomics: study of DNA sequences
Proteomics: study of proteins produced in a specific organism or cell type
Bioinformatics: using computer databases to store and analyze sequence information

15. Applications of Biotechnology Transgenic Bacteria
Transgenic: organism that contains a gene from another species in all of its cells
Transgenic bacteria that produce
Human Insulin
Human growth factor
tissue plasminogen activator (t-PA)
hepatitis B vaccine

16. Applications of Biotechnology Transgenic Plants
Bt Corn: Produces its own Pesticide
Roundup Ready Soybeans are resistant to herbicide
“Golden” rice with beta-carotene and extra iron

17. Human gene product will be released in milk
Applications of Biotechnology Transgenic Animals Make Human Gene Products
Human gene product will be released in milk

18. Same type of mouse “rescued” by blocking action of human gene
Applications of Biotechnology Transgenic Animals as Models of Human Diseases
Transgenic Mouse with defective human hormone gene interfering with growth
Same type of mouse “rescued” by blocking action of human gene

19. Ex vivo: Gene is introduced into cells outside the body
Andrew Gobea
Ex vivo: Gene is introduced into cells outside the body
Gene Therapy for SCID

20. Aerosol spray delivers gene
Cystic Fibrosis
In vivo Gene Therapy
Aerosol spray delivers gene
Injection of gene
Blindness: Leber’s Congenital Amaurosis (LCA)