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Prepare the calendar with when you are going
to do each assignment
http://www.youtube.com/watch?v=
v2T8Y3-8674
1) Check grades on AP Board. What do you need
to do to get an A or B.
2) If black body and red eyes always appears in
offspring then genes are linked/not linked.
3) Write the 5 Hardy-Weinberg conditions
4) Give 3 interpretations of this data
a) The dominant is______________
b) Sex-linked or autosomal?_________
c) Ratio_______________
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Part A—4 pts
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Part B—4pts
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Part C—4pts
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Antonio—Advanced!
Calvin, Wendy, Genesis, Jade, Diana, Kendy, Chelsea--Proficient
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1. Answer: C
2. Answer: B
3. Answer: B
4. Answer: D
5. Answer: A
6. Answer: C
7. Answer: E
8. Answer: D
9. Answer: E
10. Answer: E
11. Answer: B
12. Answer: C
13. Answer: D
14. Answer: C
15. Answer: B
16. Answer: A
17. Answer: C
18. Answer: C 19. Answer: A
20. Answer: D
21. Answer: C
22. Answer: E
23. Answer: E
24. Answer: E
25. Answer: D
26. Answer: B
27. Answer: A
28. Answer: C
29. Answer: B
30. Answer:
D
31. Answer: B
32. Answer: C
33. Answer: D
34. Answer: A
35. Answer: E
36. Answer: B
37. Answer: E
38. Answer: C
39. Answer: E
40. Answer: C
41. Answer: E
42, Answer: B
43. Answer: D
44. Answer: C
45. Answer: D
46. Answer: D
47. Answer: D
Part 1: Basic Biotechnology
TACGCACATTTACGTACGCGGATGCCGCGACTATGATC
ACATAGACATGCTGTCAGCTCTAGTAGACTAGCTGACT
human genome
CGACTAGCATGATCGATCAGCTACATGCTAGCACACYC
GTACATCGATCCTGACATCGACCTGCTCGTACATGCTA
3.2
billion
bases
CTAGCTACTGACTCATGATCCAGATCACTGAAACCCTA
GATCGGGTACCTATTACAGTACGATCATCCGATCAGAT
CATGCTAGTACATCGATCGATACTGCTACTGATCTAGC
TCAATCAAACTCTTTTTGCATCATGATACTAGACTAGC
TGACTGATCATGACTCTGATCCCGTAGATCGGGTACCT
ATTACAGTACGATCATCCGATCAGATCATGCTAGTACA
TCGATCGATACTGCTACTGATCTAGCTCAATCAAACTC
TTTTTGCATCATGATACTAGACTAGCTGACTGATCATG
ACTCTGATCCCGTAGATCGGGTACCTATTACAGTACGA
TCATCCGATCAGATCATGCTAGTACATCGATCGATACT
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Genetic Engineering
◦ manipulation of DNA
◦ if you are going to engineer DNA & genes &
organisms, then you need a
set of tools to work with
◦ this unit is a survey
of those tools…
Our tool kit…
PCR
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Bacteria review
◦ one-celled prokaryotes
◦ reproduce by mitosis
 binary fission
◦ rapid growth
 generation every ~20 minutes
 108 (100 million) colony overnight!
◦ dominant form of life on Earth
◦ incredibly diverse
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Single circular chromosome
◦ haploid
◦ naked DNA
 no histone proteins
◦ ~4 million base pairs
 ~4300 genes
 1/1000 DNA in eukaryote
How have these
little guys gotten to
be so diverse??
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Single circular chromosome
◦ haploid
◦ naked DNA
 no histone proteins
◦ ~4 million base pairs
 ~4300 genes
 1/1000 DNA in eukaryote
How have these
little guys gotten to
be so diverse??
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Small supplemental circles of DNA
 5000 - 20,000 base pairs
 self-replicating
◦ carry extra genes
 2-30 genes
 genes for antibiotic resistance
◦ can be exchanged between bacteria
 bacterial sex!!
 rapid evolution
◦ can be imported from
environment
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A way to get genes into bacteria easily
◦ insert new gene into plasmid
◦ insert plasmid into bacteria = vector
◦ bacteria now expresses new gene
 bacteria make new protein
gene from
other organism
cut DNA
plasmid
recombinant
plasmid
+
vector
glue DNA
transformed
bacteria
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Plasmids used to insert new genes into
bacteria
cut DNA
gene we
want
like what?
…insulin
…HGH
…lactase
cut plasmid DNA
Cut DNA?
DNA scissors?
ligase
recombinant
plasmid
insert “gene we want”
into plasmid...
“glue” together
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Restriction enzymes
◦ restriction endonucleases
◦ discovered in 1960s
◦ evolved in bacteria to cut up foreign DNA
 “restrict” the action of the attacking organism
 protection against viruses
& other bacteria
 bacteria protect their own DNA by methylation &
by not using the base
sequences recognized
by the enzymes
in their own DNA
Madam I’m Adam
Action of enzyme
 restriction site
CTGAATTCCG
GACTTAAGGC
◦ symmetrical “palindrome”
◦ produces protruding ends
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◦ cut DNA at specific sequences
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 sticky ends
CTG|AATTCCG
 will bind to any complementary DNA GACTTAA|GGC
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Many different enzymes
◦ named after organism they are found in
 EcoRI, HindIII, BamHI, SmaI
1960s | 1978
Werner Arber
Daniel Nathans
Restriction enzymes are
named for the organism
they come from:
EcoRI = 1st restriction
enzyme found in E. coli
Hamilton O. Smith
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Cut DNA at specific sites
◦ leave “sticky ends”
restriction enzyme cut site
GTAACGAATTCACGCTT
CATTGCTTAAGTGCGAA
restriction enzyme cut site
GTAACG AATTCACGCTT
CATTGCTTAA GTGCGAA
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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
cut sites
gene you want
cut sites
TTGTAACGAATTCTACGAATGGTTACATCGCCGAATTCACGCTT
AACATTGCTTAAGATGCTTACCAATGTAGCGGCTTAAGTGCGAA
AATTCTACGAATGGTTACATCGCCG
sticky ends GATGCTTACCAATGTAGCGGCTTAA
cut sites
isolated gene
chromosome want to add gene to
AATGGTTACTTGTAACG AATTCTACGATCGCCGATTCAACGCTT
TTACCAATGAACATTGCTTAA GATGCTAGCGGCTAAGTTGCGAA
DNA ligase joins the strands
sticky ends stick together
Recombinant DNA molecule
chromosome with new gene added
TAACGAATTCTACGAATGGTTACATCGCCGAATTCTACGATC
CATTGCTTAAGATGCTTACCAATGTAGCGGCTTAAGATGCTAGC
How can
bacteria read
human DNA?
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Gene produces protein in different
organism or different individual
human insulin gene in bacteria
TAACGAATTCTACGAATGGTTACATCGCCGAATTCTACGATC
CATTGCTTAAGATGCTTACCAATGTAGCGGCTTAAGATGCTAGC
“new” protein from organism
ex: human insulin from bacteria
aa aa aa aa aa aa aa aa aa aa
bacteria
human insulin
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Since all living
organisms…
◦ use the same DNA
◦ use the same code
book
◦ read their genes
the same way
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Transformation
◦ insert recombinant plasmid
into bacteria
◦ grow recombinant bacteria in agar cultures
 bacteria make lots of copies of plasmid
 “cloning” the plasmid
◦ production of many copies of inserted gene
◦ production of “new” protein
 transformed phenotype
DNA
 RNA  protein  trait
gene from
other organism
recombinant
plasmid
+
vector
plasmid
grow
bacteria
harvest (purify)
protein
transformed
bacteria
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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
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Word processing metaphor…
◦ cut
 restriction enzymes
◦ paste
 ligase
◦ copy
 plasmids
 bacterial transformation
 is there an easier way??
◦ find
 ????