AP Biology 12 AP Biology 12 Concept 2: Analyzing and utilizing biotechnology tools Please refer to:...
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Transcript of AP Biology 12 AP Biology 12 Concept 2: Analyzing and utilizing biotechnology tools Please refer to:...
AP Biology 12AP Biology 12
Concept 2: Analyzing and utilizing biotechnology toolsPlease refer to: Chapter 20 in Campbell
Pg 136-141 in Holtzclaw Pg 307-313 in HoltzclawPractice Questions: Campbell: #1-4 p. 405 (20.1) #1-3 p. 411 (20.2) #1-3 p. 416 (20.3) #1,4,5,6,7,8,9,10 ,11,12 p. 424-425
Holtzclaw: Questions from p. 143-150, 310, 312-313
Transgenic Organisms Transgenic Organisms This goat makes spider silk This goat makes spider silk protein in its milk! How?protein in its milk! How?
Try This!Try This!One strand of a DNA molecule has the
sequence 3′-GGATGCCCTAGGCTTGTT-5′. Which of the following is the complementary strand?
◦3′-AACAAGCCTAGGGCATCC-5′◦3′-CCTACGGGATCCGAACAA-5′◦5′-AACAAGCCTAGGGCATCC-3′◦5′-GGATGCCCTAGGCTTGTT-3
Try This!Try This!One strand of a DNA molecule has the
sequence 3′-GGATGCCCTAGGCTTGTT-5′. Which of the following is the complementary strand?
◦3′-AACAAGCCTAGGGCATCC-5′◦3′-CCTACGGGATCCGAACAA-5′◦5′-AACAAGCCTAGGGCATCC-3′◦5′-GGATGCCCTAGGCTTGTT-3
Try This!Try This!You have isolated this eukaryotic
gene and wish to express the protein it codes for in a culture of recombinant bacteria. Will you be able to produce a functioning protein with the gene as is?A. yesB. No, the exons will need to be cut out and the
introns spliced back together.C. No, the introns will need to be cut out and the
exons spliced back together.D. No, the exons will need to be cut out, the introns
translated individually, and the peptides bound together after translation.
Try This!Try This!You have isolated this eukaryotic
gene and wish to express the protein it codes for in a culture of recombinant bacteria. Will you be able to produce a functioning protein with the gene as is?A. yesB. No, the exons will need to be cut out and the
introns spliced back together.C. No, the introns will need to be cut out and
the exons spliced back together.D. No, the exons will need to be cut out, the introns
translated individually, and the peptides bound together after translation.
We’ve come a long way!We’ve come a long way!1995 – first entire genome
sequenced (bacteria)2007 – only 12 years later
◦Sequencing under way for 2000 organisms
◦Complete human genome sequenced (3 billion base pairs!)
◦Can look it up on the internet
What are other examples of What are other examples of biotechnology feats?biotechnology feats?Biotechnology: manipulation of
organism or their feats to make useful products◦Selective breeding (farms, dogs)
Choosing who breeds with who!
◦Making wine and cheese and bread Using bacteria/yeast!
◦Genetic engineering Making new proteins
So... What are the main So... What are the main techniques for manipulating techniques for manipulating DNA? DNA? Learning IntentionsLearning IntentionsYou must know:
◦The terminology of biotechnology.◦The steps in gene cloning with special
attention to the biotechnology tools that make cloning possible.
◦The key ideas that make PCR possible◦How gel electrophoresis can be used
to separate DNA fragments or protein molecules
◦Examples of genetic engineering products
Learning Intentions for AP Learning Intentions for AP Investigation Labs – See Investigation Labs – See HandoutHandout AP Investigation 7: Bacterial Transformation
◦ The principles of bacterial transformation, including how plasmids are engineered and taken up by cells
◦ Factors that affect transformation efficiency Calculate transformation efficiency and express the results in scientific notation Predict and justify how a change in the basic protocol for bacterial transformation would
affect transformation efficiency
◦ How to verify and screen for transformed cells
◦ Bacterial transformation is a type of horizontal gene transfer, and increases genetic variation
AP Investigation 9: Restriction Enzyme Analysis ◦ The function of restriction enzymes and their role in genetic
engineering
◦ How gel electrophoresis separate DNA fragments
◦ How to use a standard curve to determine the size of DNA fragments Apply mathematical routines to construct a graph of DNA fragments of known size Use this standard curve to determine the size of unknown DNA fragments Use the results of gel electrophoresis to map the restriction sites of a bacterial plasmid
Review: PCR and Review: PCR and ElectrophoresisElectrophoresis http://learn.genetics.utah.edu/content/labs/pcr/ http://www.phschool.com/science/biology_place/
labbench/lab6/intro.html http://learn.genetics.utah.edu/content/labs/gel/ Complete: Investigation – How Can Gel
Electrophoresis Be Used to Analyze DNA?
Try This!Try This! This segment of DNA is cut at restriction sites 1 This segment of DNA is cut at restriction sites 1 and 2, which creates restriction fragments A, B, and 2, which creates restriction fragments A, B, and C. and C. Which of the following electrophoretic gels Which of the following electrophoretic gels represents the separation of these fragments?represents the separation of these fragments?
a)
b)
c)
d)
Let’s learn how to “clone” Let’s learn how to “clone” genes.genes.Gene cloning:
◦Process of producing multiple copies of specific DNA segments, making recombinant DNA in the process.
◦Tools: Restriction enzymes (Campbell – some one log into their
student account please to show the class) – discovered in the 1960s by bacterial researchers
Cloning GenesCloning GenesRestriction enzymes
◦How do they help bacteria in the wild? Cut up foreign DNA (ex: phage DNA)
◦How do they work? Recognize restriction sites on DNA
Symmetrical/palindromal 4-8 nuclotide base pairs long
Make many cuts – produce restriction fragments
Cuts in a reproducible way Bacterial DNA protected by methyl groups on
restriction sites.
Try This!Try This!Which of the following DNA
molecules is most likely to be cut by a restriction enzyme?A. 5′-AAGCCT-3′B. 5′-GGAAGG-3′C. 5′-GGATCC-3′D. 5′-AATTAA-3′
Try This!Try This!Which of the following DNA
molecules is most likely to be cut by a restriction enzyme?A. 5′-AAGCCT-3′B. 5′-GGAAGG-3′C. 5′-GGATCC-3′ (palindrome)D. 5′-AATTAA-3′
The process of Cloning The process of Cloning genesgenesCloning a Gene in Bacteria Campbell
Activity!
NOW… Transformation NOW… Transformation Lab!Lab!Handout – LabPrepare by reading and making flow
chart.
INTENSE PROCEDURE!!!◦Make sure you understand it. Come into
the lab knowing what you are doing. If you have questions, please ask!.
◦Thurs, Jan 29th ◦Fri, Jan 30th – lunch: Results
Also…Also…You should now be able to answer all the questions of this section…
Questions…