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Nekeisha Nekeisha

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Nekeisha - PPT Presentation

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

vector dna hpv vaccine dna vector vaccine hpv restriction recombinant insert cut enzymes students script order infected development steps

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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

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