1. Introduction to Yeast Plasmids
Doug Wassarman
PUBS
Sept. 19, 2017

2. What is a plasmid? Small DNA molecule Independent replication Circular
Used for horizontal gene transfer
Contain genes for drug resistance, virulence factors, etc.
Chromosomal DNA
Plasmids

3. What do we use plasmids for?
Cloning – DNA manipulation
khanacademy.org
addgene.org

4. What do we use plasmids for?
Cloning – DNA manipulation
Protein Expression
khanacademy.org

5. Elements of a plasmid

6. Elements of a plasmid
Replication origin

7. Replication Origin Integrating Episomal Chromosomal DNA Plasmids ori

8. Elements of a plasmid
Replication origin

9. Elements of a plasmid
Replication origin
Selectable Marker

10. Antibiotic Resistance
Selectable Marker
Antibiotic Resistance

11. Antibiotic Resistance
Selectable Marker
Antibiotic Resistance
Auxotrophy
-Ura Medium

12. Antibiotic Resistance
Selectable Marker
Antibiotic Resistance
Auxotrophy
-Ura Medium

13. Elements of a plasmid
Replication origin
Selectable Marker

14. Elements of a plasmid Replication origin Selectable Marker
Gene of Interest

15. Elements of a plasmid Replication origin Selectable Marker
Gene of Interest
Promoter

16. Promoters
Constitutive
Always “on”
Selectable markers

17. Promoters Always “on” Selectable markers “Off” by default
Constitutive
Inducible
Always “on”
Selectable markers
“Off” by default
Can be turned on
Glucose
off
Galactose on

18. Elements of a plasmid Replication origin Selectable Marker
Gene of Interest
Promoter

19. Questions?