Download Biotechnology2

Biotechnology
AP Biology
2007-2008
D.N.A
 Review HW concept questions and be
ready to ask any questions that you
may have
 Take 15 minutes to COMPLETE building
your protein synthesis poster

AP Biology
I will be coming around for you to
discuss your poster with me
A Brave New World
AP Biology
TACGCACATTTACGTACGCGGATGCCGCGACTATGATC
ACATAGACATGCTGTCAGCTCTAGTAGACTAGCTGACT
human genome
CGACTAGCATGATCGATCAGCTACATGCTAGCACACYC
GTACATCGATCCTGACATCGACCTGCTCGTACATGCTA
3.2
billion
bases
CTAGCTACTGACTCATGATCCAGATCACTGAAACCCTA
GATCGGGTACCTATTACAGTACGATCATCCGATCAGAT
CATGCTAGTACATCGATCGATACTGCTACTGATCTAGC
TCAATCAAACTCTTTTTGCATCATGATACTAGACTAGC
TGACTGATCATGACTCTGATCCCGTAGATCGGGTACCT
ATTACAGTACGATCATCCGATCAGATCATGCTAGTACA
TCGATCGATACTGCTACTGATCTAGCTCAATCAAACTC
TTTTTGCATCATGATACTAGACTAGCTGACTGATCATG
ACTCTGATCCCGTAGATCGGGTACCTATTACAGTACGA
TCATCCGATCAGATCATGCTAGTACATCGATCGATACT
AP Biology
Biotechnology today
 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…

AP Biology
Our tool kit…
Bacteria
 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

AP Biology
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


mix heat-killed
pathogenic &
non-pathogenic
bacteria
import bits of chromosomes from
other bacteria
incorporate the DNA bits into their
own chromosome
 express new genes
 transformation
 form of recombination
AP Biology
mice die
Plasmids
 Small supplemental circles of DNA
 5000 - 20,000 base pairs
 self-replicating

carry extra genes
 2-30 genes

can be exchanged between bacteria
 bacterial sex!!
 rapid evolution

can be imported from
environment
AP Biology
How can plasmids help us?
 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
AP Biology
recombinant
plasmid
+
vector
glue DNA
transformed
bacteria
Biotechnology
 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
APplasmid
Biology
insert “gene we want”
into plasmid...
“glue” together
How do we cut DNA?
 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
AP Biology
cut DNA at specific sequences CTGAATTCCG
 restriction site

produces protruding ends
GACTTAAGGC



Restriction enzymes
 Action of enzyme
 sticky ends
CTG|AATTCCG
 will bind to any complementary DNA
GACTTAA|GGC
 Many different enzymes

named after organism they are found in
 EcoRI, HindIII, BamHI, SmaI
AP Biology
Restriction enzymes
 Cut DNA at specific sites

leave “sticky ends”
restriction enzyme cut site
GTAACGAATTCACGCTT
CATTGCTTAAGTGCGAA
restriction enzyme cut site
GTAACG AATTCACGCTT
CATTGCTTAA GTGCGAA
AP Biology
Sticky ends
 Cut other DNA with same enzymes


leave “sticky ends” on both
can glue DNA together at “sticky ends”
GTAACG AATTCACGCTT
CATTGCTTAA GTGCGAA
AP Biology
gene
you want
GGACCTG AATTCCGGATA
CCTGGACTTAA GGCCTAT
chromosome
want to add
gene to
GGACCTG AATTCACGCTT
CCTGGACTTAA GTGCGAA
combined
DNA
Sticky ends help glue genes together
cut sites
gene you want
cut sites
TTGTAACGAATTCTACGAATGGTTACATCGCCGAATTCACGCTT
AACATTGCTTAAGATGCTTACCAATGTAGCGGCTTAAGTGCGAA
AATTCTACGAATGGTTACATCGCCG
GATGCTTACCAATGTAGCGGCTTAA
sticky ends
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
AP Biology
CATTGCTTAAGATGCTTACCAATGTAGCGGCTTAAGATGCTAGC
How can
bacteria read
human DNA?
Why mix genes together?
 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
AP Biology
human insulin
Transforming Bacteria
The gene for HGH
(growth hormone) is
removed from a
human cell
AP Biology
Plasmid
The plasmid is cut at certain
sequences in the DNA using the same
Restriction Enzyme used to cut the
HGH gene from a human cell
The plasmid will have sticky ends in
addition to the HGH gene
AP Biology
The gene for HGH is
then mixed with the
bacterial plasmid,
and the HGH is
incorporated into the
plasmid
DNA Ligase
covalently bond the
sugar phosphate
backbone
The plasmid is then
added to the bacteria
AP Biology
The bacteria now has the
gene for HGH, and has the
ability to produce it
This shows the process of
transformation in bacteria
AP Biology
The code is universal
 Since all living
organisms…



AP Biology
use the same DNA
use the same code
book
read their genes
the same way
Copy (& Read) DNA
 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
AP Biology
Green with envy??
Jelly fish “GFP”
AP Biology
Transformed vertebrates
 HW: Read 12.10-12.15
 Complete Recombinant (Plasmid) DNA
questions
AP Biology
AP Biology
Cut, Paste, Copy, Find…
 Word processing metaphor…

cut
 restriction enzymes

paste
 ligase

copy
 plasmids
 bacterial transformation
 is there an easier way??

find
 ????
AP Biology
D.N.A
Objective: SWBAT examine the arguments
supporting and opposing the use of Golden Rice
and evaluate the merits of using GMO’s in society.
 Explain how DNA segments can be cut
and spliced together to produce
recombinant DNA. How do the
segments “find” each other and stick
together? How is recombinant DNA
then cloned to produce multiple copies
of the gene?
AP Biology
 A restriction enzyme is used to cut DNA




at a specific nucleotide sequence
It cuts the two strands of DNA, forming
“sticky ends” – can easily hydrogen
bond to other DNA strands
Enzyme DNA ligase links pieces of DNA
A fragment of DNA can be spliced into
a plasmid (circular bacterial DNA)
Transformation – bacteria can take up
the plasmid and begin to replicate the
newly DNA sequence
AP Biology
GOLDEN RICE DEBATE
 Ethical Question: Should Golden Rice
be used in developing nations to
combat vitamin A deficiency
AP Biology
 STEP 1: RESEARCH
GET INTO TEAMS OF 4
 SOME TEAMS WILL RESEARCH PRO, OTHER CONS
 MAKE SURE YOU TAKE NOTES
 YOU WILL NOT BE ABLE TO USE YOUR RESEARCH
PAPER (1O MIN)
 STEP 2: CONVINCE OTHERS OF YOUR POSITION
 CREATE A MINI TEAM (2) WITHIN YOUR GROUP OF 4
 COMBINE WITH ANOTHER MINI TEAM AND PREPARE
YOUR ARGUMENT (5 MIN FOR PRO, 5 MIN FOR CON)
 STEP 3: CONVINCE OTHERS OF THE OPPOSING POSITION
 FIND A MINI TEAM THAT HAD AN OPPOSING VIEW
 NOW YOU MUST ARGUE FOR THAT OPPOSING VIEW (5
MIN CON, 5 MIN PRO)

AP Biology
AP Biology