Team:NYMU-Taipei/Experiment/Wet Lab

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Revision as of 18:49, 27 September 2013 by S10101054 (Talk | contribs)

Data for Our Favorite Parts 1. A ‘’’ OxyR-induced Promoter’’’ BBa_K1104201(TrxC promotor) 2. A ‘’’Transcription Factor Activated by Oxidative Stress’’’ BBa_K1104200 (OxyR ) we apply a constitutive promoter to the OxyR coding sequence to overexpress the OxyR protein, which is expected to boost the sensitivity of our ROS-induced promotors. 3. An ‘’’antimicrobial peptide’’’ BBa_K1104200 (Defensin1 ) the mechanism which insects use to protect against the invasion of microbes.


Data for pre-Existing and Optimized Parts We successfully improved the biobrick part: ahpC promoter ahpC (K362001). Originally created by the KIT-Kyoto 2010 iGEM team by taking the upstream 1000bp of the AhpC gene, we improved the promoter by first mutating the PstI cutting site to make it conform to Assembly standard 10. We then designed specific primers to cut short the original sequence of AhpCp promoter to trim out unneeded sequences make it more efficient. More details about the design can be found on the Team:NYMU-Taipei/Project/Inhibition/Sensor Sensor page.

The supplementary characterization and the application of AhpC promoter has been added to the experience page of BBa_K1104203 (AhpCp1000).

The AhpCp1000 derived OxyR-induced promoters we created are as follows:





National Yang Ming University


Contents

Inhibition of Spore Germination Testing---Mannosidase Function Assay

Nymu 流程圖.png

We define the germination degree as follows:

1 point:A spore begins its budding reproduction.
2 points:A new spore is ccompletely germinanated from the old one.
3 points:A cluster of duplicated spores is formed.
4 points:A Spore extrude a polar filament.

And so far we have accomplished experiments of the first two groups, discovering that spores underwent the germination procesure got much more points than the control group. Next we will compare the points of group 3 and group 4 to see if Bee. coli can inhibit the germination of N. ceranae.

N. ceranae Experiment Protocols

Extraction of N. ceranae Spores from Bees

Extraction.png

Purification of N. ceranae Spores

Nymu-Purification.png

N. ceranae Genomic DNA Extraction

Nymu-Genomic.png

Germination of N. ceranae Spores

Nymu-Germination.PNG

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