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…