Download ppt

AP Biology 12
Concept 2: Analyzing and
utilizing biotechnology tools
Please refer to:
Chapter 20 in Campbell
Pg 136-141 in Holtzclaw
Pg 307-313 in Holtzclaw
Practice 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
This goat makes spider silk protein
in its milk! How?
Try This!
One strand of a DNA molecule has the
sequence 3′-GGATGCCCTAGGCTTGTT-5′.
Which of the following is the complementary
strand?
A.3′-AACAAGCCTAGGGCATCC-5′
B.3′-CCTACGGGATCCGAACAA-5′
C.5′-AACAAGCCTAGGGCATCC-3′
D.5′-GGATGCCCTAGGCTTGTT-3
Try This!
One strand of a DNA molecule has the
sequence 3′-GGATGCCCTAGGCTTGTT-5′.
Which of the following is the complementary
strand?
A.3′-AACAAGCCTAGGGCATCC-5′
B.3′-CCTACGGGATCCGAACAA-5′
C.5′-AACAAGCCTAGGGCATCC-3′
D.5′-GGATGCCCTAGGCTTGTT-3
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. yes
B. 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!

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. yes
B. 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!
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
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 techniques
for manipulating DNA?
Learning Intentions

You 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
Investigation Labs – See Handout

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 Electrophoresis




http://learn.genetics.utah.edu/content/labs/pcr/
http://www.phschool.com/science/biology_place/labbench/lab6/intr
o.html
http://learn.genetics.utah.edu/content/labs/gel/
Complete: Investigation – How Can Gel Electrophoresis Be Used
to Analyze DNA?
Try This!
This segment of DNA is cut at restriction sites 1 and 2, which
creates restriction fragments A, B, and C.
Which of the following electrophoretic gels represents the
separation of these fragments?
a)
b)
c)
d)
Let’s learn how to “clone” 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 Genes

Restriction 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!

Which of the following DNA molecules is
most likely to be cut by a restriction
enzyme?
A.
B.
C.
D.
5′-AAGCCT-3′
5′-GGAAGG-3′
5′-GGATCC-3′
5′-AATTAA-3′
Try This!

Which of the following DNA molecules is
most likely to be cut by a restriction
enzyme?
A.
B.
C.
D.
5′-AAGCCT-3′
5′-GGAAGG-3′
5′-GGATCC-3′ (palindrome)
5′-AATTAA-3′
The process of Cloning genes

Cloning a Gene in Bacteria
Campbell Activity!
NOW… Transformation Lab!
Handout – Lab
 Prepare 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…

You should now be able to answer all
the questions of this section…
Questions…