Nogal Gabriel Pineda Keith Neeves Judy Schoonmaker Delphi Chatterjee Diane Ordway Erika Tinoco Goals and Outcomes Unit Engineering recombinant DNA vaccines Goal To understand the molecular biology sequence of antigen ID: 382185
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Slide1
Nekeisha
Nogal
, Gabriel Pineda,
Keith Neeves, Judy
Schoonmaker
,
Delphi
Chatterjee
, Diane Ordway,
Erika
TinocoSlide2
Goals and Outcomes
Unit
: Engineering recombinant DNA vaccines
Goal: To understand the molecular biology sequence of antigen
vaccine
Goals for Today:
Making a recombinant DNA vector
Students will understand how restriction enzymes work
Students will understand how DNA ligase works
Outcomes
Students will be able to
organize
the main steps in DNA vaccine
development
Students
will be able to
recognize
the specificity of
ligation
sites following cleavage by restriction
enzymes
Students will be able to
create
a script that
applies
the concepts of gene insertionSlide3
Course Context
Biology I or II
Freshmen
~100 students
Prior
knowledge
DNA structure
Central dogma
Restriction enzymes
Vaccines and immune responseSlide4
DNA Vaccine
Recombinant Vector
Have you gotten
YOUR
HPV Vaccine?
HPV Vaccine
20 million Americans currently infected
6 million infected/
yr
50% sexually active people have HPV at some point
http://hplusmagazine.com/2009/08/31/take-shot/
Accessed 8/3/12Slide5
Steps in DNA vaccine development
Identify HPV immunogenic protein
Choose a vector
Culture transformed bacterial host cells
Purify recombinant vector
Deliver recombinant HPV vector vaccine
Determine coding DNA for insert
Cut vector and DNA insert with restriction enzymes
Ligate DNA insert into vectorSlide6
Steps in DNA vaccine development
Identify HPV immunogenic protein
Choose a vector
Culture transformed bacterial host cells
Purify recombinant vector
Deliver recombinant HPV vector vaccine
Determine coding DNA for insert
Cut vector and DNA insert with restriction enzymes
Ligate DNA insert into vectorSlide7
Campbell Essential Biology with Physiology, Simon et al., 3
rd
Ed.
Cut vector and DNA insert with restriction enzymesSlide8
The restriction enzymes
RE1
and
RE2
cut along the red lines:
If one sample of DNA was cut with RE1 and another with RE2, and these two samples were mixed and treated with DNA ligase, what would happen?
A. DNA cut with RE1 and RE2 would be ligated (joined together)
with no preference.
B. The RE1-cut DNA would be ligated to itself.
C. The RE2-cut DNA would be ligated to itself.
D. Both B and C.
E. No DNAs would be ligated.Slide9
Write a script for this
movie
using these terms:
Restriction enzyme
Plasmid
DNA ligase
DNA insert
Sticky ends
Restriction site
Peer review script
Turn in revised script to teacher.
http://www.youtube.com/watch?v=aA5fyWJh5S0
Ligate HPV specific DNA insert into vectorSlide10
DNA Vaccine
Recombinant Vector
Have you gotten
YOUR
HPV Vaccine?
HPV Vaccine
20 million Americans currently infected
6 million infected/
yr
50% sexually active people have HPV at some point
http://hplusmagazine.com/2009/08/31/take-shot/
Accessed 8/3/12Slide11
Homework Assignment(summative assessment)
Make a concept map that shows how a scientist would proceed from a gene sequence for a protein to a vaccine using recombinant technology.
You should use the terms below as a skeleton framework. Add as many additional terms as you feel appropriate.
Restriction enzyme
Vector
DNA ligase
DNA insert
Sticky ends
Vaccine Slide12
If you had an HPV-specific DNA insert that has sticky ends created by digestion with Bam
HI, which of the three restriction enzymes below could be used to cut a vector to create a recombinant vector vaccine for HPV?
Bam
HI
only
Bam
HI
and
Bsl
I
BslI and EcoRI
EcoRI
onlyEcoRI and BamHI
Exam Question(summative assessment)BamHI – 5’-G|GATCCBslI – 5’-C|GATCGEcoRI – 5’-G|AATTCSlide13
Alignment
Learning Objective
Assessment
Active learning
Low Order/
High Order
Students will be able to
organize
the main steps in DNA vaccine development
Formative: Self-check based on key
Strip sequence
Low Order
Bloom 3
Students will be able to
recognize
the specificity of
ligation sites following cleavage by restriction enzymes
Formative: Clicker question-two polls
Clicker question + peer instruction
High Order
Bloom 3-4
Students will be able to
create
a script that
synthesizes
the key concepts of gene insertion
Formative: Peer evaluation
Summative: Grade the script
Writing script of movie
High Order
Bloom 5Slide14
Diversity
Different learning styles:
Kinesthetic – strip sequence
Visual – movie
Writing – script
Auditory – movie sounds
Different personalities:
Activities that appeal to introverts and extroverts