Transcript Creating a lipid biosynthesis hybrid: E. nemone

Creating a lipid biosynthesis hybrid:
E. nemone
Teresa and Jodi
Goals
• Combine our research interests
– Teresa: lipid biosynthesis
– Jodi: coral symbiosis and genomics
• Teach our students
• Try something new
• Keep it low risk
Combine our research interests:
Roles of Lipids in
Coral Symbiosis and Bleaching
Lipids involved in
establishment
of symbiosis
Symbiotic unit
Photosynthetic
symbionts
Anemone host
Lipids produced during
thermal stress
and loss of symbiosis
+
E-nemone!
Lipid Biosynthetic Pathways
PE
PC
PA
PS
CDP-DAG
PI
PG-P
PG
E. coli pathway
Eukaryotic pathway
CL
Eukaryotic only lipids
Teach our students:
A hybrid course to build E. nemone
• Combine our two existing courses
– Chem 145: Chemistry Research Techniques
• for accelerated freshmen
• 9 students
– Biol 244: Genomics
• sophomore/junior lab course
• 16 students
• Six week intersection
Research questions:
• Can we alter the lipidome of E. coli?
• Can we learn about Anemone lipid biosynthesis and function?
Week 1 Intro to project
Context of project
Brainstorming about genomics and biochemistry approaches
Learn about E. coli and eukaryotic lipid biosynthesis pathways
Week 2 Identify target genes in E. coli and anemone genomes
Choose target enzymes
Chem students search for E. coli genes
Bio students search for genes in anemone genomes
Week 3 Decide what is feasible to engineer and get started
Presentation of genes/discussion of which genes to target
Synthetic Bio (Bio Bricks) discussion
Design primers
Week 4 Build the hybrid E. nemone:
PCR, ligation, transformations, colony PCR
Week 5 Characterize lipids:
Grow cells, extract lipids, TLC, MS
Week 6 Presentations
Discussion: what would we do next?
Synthetic Biology Approach
We can use parts from the mini-registry
pLac --- RBS --- RFP:GOI --- tt
pLac --- RBS --- GOI --- RFP --- tt
pLac --- RBS --- GOI --- tt
GOI = Gene of Interest
RFP = Red Fluorescent Protein
Outcomes
• Chem students learn genomics
• Genomics students learn biochemistry
• Development of interdisciplinary
collaborations
• Introduce Synthetic Biology to Vassar
• Develop tools to enhance scholarship
• Engage students in REAL research!