Results

Inoculations of GFP Constructs (58 tubes)

1.P-Lambda-R--LacI--GFP (no deg tag)

1. 3 tubes in each cell type (DH10, Keio Wild, Keio ClpP) induced with 0 uM IPTG (plain LB)
2. 3 tubes of each cell type (DH10, Keio Wild, Keio ClpP) induced with 100 uM IPTG (plain LB)
3. RESULTS:

2. P-Lambda-R--LacI--GFP (DAS)

1. 3 tubes in each cell type (DH10, Keio Wild, Keio ClpP) induced with 0 uM IPTG (plain LB)
2. 3 tubes of each cell type (DH10, Keio Wild, Keio ClpP) induced with 100 uM IPTG (plain LB)
3. RESULTS:

3. P-Lambda-R--LacI--GFP (LAA)

1. 3 tubes in each cell type (DH10, Keio Wild, Keio ClpP) induced with 0 uM IPTG (plain LB)
2. 3 tubes of each cell type (DH10, Keio Wild, Keio ClpP) induced with 100 uM IPTG (plain LB)
3. RESULTS:

4. Plain LB, DH10 cells in plain LB, Keio Wild cells in plain LB, and Keio ClpP cells in plain LB
5. RESULTS:

1. 10/17: The GFP constructs were brought to the plate reader at Georgia Tech. Although cells were grown in the liquid culture, they did not fluoresce.

6. Pictures:

Data from the Plate Reader

All of the GFP constructs as liquid cultures.

Julia Leveille and Lauren Hong with the 58 liquid cultures before analyzing them on the plate reader at Georgia Tech.

Troubleshooting for Lack of Proper Tube Labeling

We plated our constructs in all three cell types(DH10, Keio Wild, and Keio ClpP) on Kanamycin, Tetracycline, Chloramphenicol, and Ampicillin. We were testing to verify what backbones the plasmids were in. As the image shows, our cells grew in most of the plates resistant to the specific antibiotics. We are in the process of figuring out how we obtained these results, but we hypothesized that our cells contain constructs with all the vectors with backbones resistant to those antibiotics.

Sequencing Results

The R0011 ClpX part of our DNA construct was sent in for sequencing and confirmed to be matching DNA through Eurofins MWG Operon.


The ClpP B0033 CI part of our DNA construct was also sent in for sequencing and confirmed to be matching DNA through Eufofins MWG Operon.

Expected Results

Medals

Bronze Team formed and registered
Wiki completed
Poster completed
Presentation ready for Jamboree
Attributed all work done for project
Part (BBa_K1911001) documented and submitted
All forms submitted
Silver New part (BBa_K1911000) documented, submitted, and validated
Collaboration: iGEM teams of Alverno California and CAPS_Kansas and Georgia State University (GSU), the Styczynski Lab from the Georgia Institute of Technology (GaTech), Centers for Disease Control and Prevention (CDC), New England Biolabs, and Lambert High School’s engineering department
Human Practices: Maker Faire, Atlanta Science Festival (ASF), Regional Education Service Agencies (RESA), Next Generation Focus (NGF), Congenital Heart Defect Walk, Sharon Science Days, Micronutrient Discussion with the CDC, collaboration with the Styczynski Lab from GaTech
Gold
Integrated Human Practices: collaboration with Styczynski Lab at GaTech with real-world applications of improving the zinc biosensor for malnourished and micronutrient-deficient countries
Improved previous iGEM project parts: BBa_K1033906, BBa_M0050-LAA, BBa_M0052-DAS
Proof of Concept: GFP analysis