Introduction to Yeast Plasmids

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Presentation transcript:

Introduction to Yeast Plasmids 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. Chromosomal DNA Plasmids https://en.wikipedia.org/wiki/File:Plasmid_(english).svg

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

What do we use plasmids for? Cloning – DNA manipulation Protein Expression khanacademy.org https://www.novoprolabs.com/novostyle/images/protein-expression-timeline.jpg

Elements of a plasmid

Elements of a plasmid Replication origin

Replication Origin Integrating Episomal Chromosomal DNA Plasmids ori https://en.wikipedia.org/wiki/File:Plasmid_(english).svg

Elements of a plasmid Replication origin

Elements of a plasmid Replication origin Selectable Marker

Antibiotic Resistance Selectable Marker Antibiotic Resistance http://biosiva.50webs.org/dna%20cl10.gif

Antibiotic Resistance Selectable Marker Antibiotic Resistance Auxotrophy -Ura Medium http://biosiva.50webs.org/dna%20cl10.gif

Antibiotic Resistance Selectable Marker Antibiotic Resistance Auxotrophy -Ura Medium http://biosiva.50webs.org/dna%20cl10.gif

Elements of a plasmid Replication origin Selectable Marker

Elements of a plasmid Replication origin Selectable Marker Gene of Interest

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

Promoters Constitutive Always “on” Selectable markers

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

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

Questions?