1. Biotechnology

2. DNA Technology
Recombinant DNA- DNA sequences from two different sources
Genetic engineering- artificial manipulation of the genetic material of organisms, including the creation of novel genetic material
Biotechnology is the development of new biological products

3. A Brave New World

4. Biotechnology today Genetic Engineering Our tool kit…
manipulation of DNA
if you are going to engineer DNA & genes & organisms, then you need a set of tools to work with
this topic is a survey of those tools…
Our tool kit…

5. 1. Bacteria Bacteria review one-celled prokaryotes
reproduce by binary fission
generation every ~20 minutes
108 (100 million) colony overnight!
dominant form of life on Earth
incredibly diverse

6. How have these little guys gotten to be so diverse??
2. Bacterial genome
Single circular chromosome
naked DNA
no histone proteins
~4300 genes
1/1000 DNA in eukaryote
Genome = all the DNA of an organism
Eukaryotes
• 1000 times more DNA
• only 10 times more genes
- introns, spacers, inefficiency
How have these little guys gotten to be so diverse??

7. 3. Transformation Bacteria are opportunists
pick up naked foreign DNA wherever it may be hanging out
have surface transport proteins that are specialized for the uptake of naked DNA
incorporate the DNA bits into their own chromosome
mix heat-killed pathogenic & non-pathogenic
bacteria
While E. coli lacks this specialized mechanism, it can be induced to take up small pieces of DNA if cultured in a medium with a relatively high concentration of calcium ions.
In biotechnology, this technique has been used to introduce foreign DNA into E. coli.
mice die

8. 4. Plasmids Small supplemental circles of DNA carry extra genes
,000 base pairs
self-replicating
carry extra genes
2-30 genes
genes for antibiotic resistance
can be exchanged between bacteria
rapid evolution
can be imported from environment
Mini-chromosomes

9. How can plasmids help us?
insert new gene into plasmid
insert plasmid into bacteria = vector
bacteria now expresses new gene
bacteria make new protein
transformed bacteria
gene from other organism
recombinant plasmid
vector
plasmid
cut DNA +
glue DNA

10.  5. How do we cut DNA? Restriction enzymes restriction endonucleases
evolved in bacteria to cut up foreign DNA
“restricts” the action of the attacking organism
protection against viruses & other bacteria
bacteria protect their own DNA by methylation 

11. Restriction enzymes Madam I’m Adam Action of enzyme
cut DNA at specific sequences
restriction site
symmetrical “palindrome”
produces protruding ends
sticky ends
will bind to any complementary DNA
Many different enzymes
named after organism they are found in
EcoRI, HindIII, BamHI, SmaI 
CTGAATTCCG
GACTTAAGGC 
CTG|AATTCCG
GACTTAA|GGC

12. chromosome want to add gene to
6. Sticky ends
Cut other DNA with same enzymes
leave “sticky ends” on both
can glue DNA together at “sticky ends”
GTAACG AATTCACGCTT
CATTGCTTAA GTGCGAA
gene you want
GGACCTG AATTCCGGATA
CCTGGACTTAA GGCCTAT
chromosome want to add gene to
GGACCTG AATTCACGCTT
CCTGGACTTAA GTGCGAA
combined DNA

13. 7. Cloning Genes Fast! PCR polymerase chain reaction
alternative method for cloning a gene
used primarily when the source of DNA is scanty or impure
applications:
ancient DNA
amplify DNA from fingerprints or
tiny amounts of blood, tissue, or
semen found at crime scenes
amplify DNA from embryonic cells
to test for genetic disorders

14. 8. Gel Electrophoresis-find the gene

15. Loading the Gel

16. Run Gel

17. Applications of Biotechnology
Forensics
Environmental use- oil- eating bacteria
Gene Therapy- replace damaged cells with healthy ones
Transgenic Animals
-Pharm Animals- have animals produce human products
Genetically Modified Organisms- more meat, disease and pest resistant etc

18. Blood from defendant’s
blood (D)
Blood from defendant’s
clothes
Victim’s
blood (V)
LE 20-17
DNA Blueprints are bands of RFLP markers
Looks like defendant is heading for jail- why?
What blood evidence likely convicted him/her?

19. Environmental Uses
Genetically microbes can extract heavy metals to clean up toxic wastes in mining
Bacteria can degrade oil in oil spills
Bacteria can degrade plastics

20. Transgenic Organisms Desired gene is implanted in fertilized egg
“Pharm” animals can produce large amounts of desired gene

21. Examples of Trangenic Animals
E. coli bacteria producing insulin
Spider silk in goat’s milk
Knock Out mice to study human disease
GFP ( green flourescent protein) as a marker

22. Uses of genetic engineering
Genetically modified organisms (GMO)
enabling plants to produce new proteins
Protect crops from insects: BT corn
corn produces a bacterial toxin that kills corn borer (caterpillar pest of corn)
Extend growing season: fishberries
strawberries with an anti-freezing gene from flounder
Improve quality of food: golden rice
rice producing vitamin A improves nutritional value
For example, a transgenic rice plant has been developed that produces yellow grains containing beta-carotene.
Humans use beta-carotene to make vitamin A.
Currently, 70% of children under the age of 5 in Southeast Asia are deficient in vitamin A, leading to vision impairment and increased disease rates.

23. Regular zebras
Glo- fish zebras

24. Jelly fish GFP-green fluorescent Transformed vertebrates
protein markers
Green with envy??
Transformed vertebrates