Collaboration

This year, we collaborated with a collegiate team NCHU-Taichung iGEM 2018 from National Chung Hsing University, Taiwan. NCHU is just located nearby Mingdao High School (15 minutes distance by car). At very beginning of the journey in iGEM 2018, we met up with each other at NCHU and Mingdao’s laboratory and school campus. We helped them host the 6th Asian-Pacific Conference hold in August. We shared with each other many ideas and dealt with problems met in the project and experiments. In this moment, just several weeks before iGEM Jamboree, we’d like to say, “Thank you, Friends. Happy to meet you because of iGEM”

We helped iGEM team NCHU to verify the bacterial cell growth affected by their BioBrick devices. They want to utilize synthetic biology to recover an environment polluted with dioxin. They created BioBricks of HAD, TonB and TLCC. They prepared the transformed E. coli with BioBricks and growth media (KB medium). Go to our lab in Mingdao, we measured the OD values of the E. coli cultured overnight and added IPTG for the induction of the gene expression and completed the assay following their protocol and instructions.

EXPERIMENT BY MINGDAO

Measure OD600 values for E. coli with HAD, TonB or TLCC genes
Dilute to OD600=0.05 with KB medium
Culture at for 2-3 hours until OD values reach approximately 0.6
Add 1mM IPTG for gene induction
Set a kinetic mode for analysis in Microplate Reader (BioTeK Synergy H1)
Analyze the data with each group for 3 repeats and report to Team NCHU

RESULT

The figure showed TonB helped bacterial growth and further improved by HAD. However, TLCC seems some toxic effect on bacterial growth.

On the other hand, we want to demonstrate our pathogen testing system in real adult mosquitoes and understand the GFP reporter system driven by GAM1 promoter can be triggered by E. coli. We prepared the plasmids of GAM1-GFP-polyA /pSB1C3 and heat-killed E. coli. Team NCUH brought us to the Mosquito Lab at Entomology Department of National Chung Hsing University and helped us microinjected DNAs into the midgut of Aedes aegypti.

EXPERIMENT BY NCHU

Make microinjection needles by pulling glass capillary tubes
Anesthetize the adult mosquitoes on ice
Fill the needle with the DNAs in Cellfectin II reagent with E. coli or not
Inject the DNA by inserting into the thorax of the mosquito.
Recover the mosquito for 2 days

RESULT

The mosquito injected with DNA plus E. coli showed fluorescence signal in Gel Imaging System and Blue LED Box after 2 days of DNA microinjection.