Doug Wassarman PUBS Sept 19 2017 What is a plasmid Small DNA molecule Independent replication Circular Used for horizontal gene transfer Contain genes for drug resistance virulence factors etc ID: 633282
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Slide1
Introduction to Yeast Plasmids
Doug Wassarman
PUBS
Sept. 19, 2017Slide2
What is a plasmid?
Small DNA molecule
Independent replication
CircularUsed for horizontal gene transferContain genes for drug resistance, virulence factors, etc.
Chromosomal DNA
Plasmids
https://
en.wikipedia.org
/wiki/
File:Plasmid
_(
english
).
svgSlide3
What do we use plasmids for?
k
hanacademy.org
a
ddgene.org
Cloning
–
DNA manipulationSlide4
What do we use plasmids for?
k
hanacademy.org
https://
www.novoprolabs.com
/
novostyle/images/protein-expression-timeline.jpg
Cloning
–
DNA manipulation
Protein ExpressionSlide5
Elements of a plasmidSlide6
Elements of a plasmid
Replication originSlide7
Replication Origin
Integrating
Episomal
ori
Chromosomal DNA
Plasmids
https://
en.wikipedia.org
/wiki/
File:Plasmid
_(
english
).
svgSlide8
Elements of a plasmid
Replication originSlide9
Elements of a plasmid
Replication origin
Selectable MarkerSlide10
Selectable Marker
Antibiotic Resistance
http://biosiva.50webs.org/dna%20cl10.gifSlide11
Selectable Marker
Antibiotic Resistance
Auxotrophy
http://biosiva.50webs.org/dna%20cl10.gif
-
Ura
MediumSlide12
Selectable Marker
Antibiotic Resistance
Auxotrophy
http://biosiva.50webs.org/dna%20cl10.gif
-
Ura
MediumSlide13
Elements of a plasmid
Replication origin
Selectable MarkerSlide14
Elements of a plasmid
Replication origin
Selectable Marker
Gene of InterestSlide15
Elements of a plasmid
Replication origin
Selectable Marker
Gene of Interest
PromoterSlide16
Promoters
Constitutive
Always “on”
Selectable markersSlide17
Promoters
Constitutive
Always “on”
Selectable markers
Inducible
“Off” by defaultCan be turned on
Glucose
Galactose
off
onSlide18
Elements of a plasmid
Replication origin
Selectable Marker
Gene of Interest
PromoterSlide19
Questions?