Team:NYMU-Taipei/Experiment/Wet Lab

From 2013.igem.org

National Yang Ming University

1 Data for Our Favorite Parts
2 Data for Pre-Existing and Optimized Parts
- 2.1 AhpCp
- 2.2 SufAp
3 List of Our Parts

Data for Our Favorite Parts

An OxyR-induced Promoter BBa_K1104201 (TrxC promoter)
A Transcription Factor Activated by Oxidative Stress BBa_K1104200 (OxyR) - We apply a constitutive promoter to the upstream region of OxyR coding sequence to overexpress the OxyR protein, which is expected to boost the sensitivity of our ROS-induced promotors.
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

AhpCp

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 Nosema Sensor page.

AhpCp1000Part:BBa_K1104203
We succesfully mutated the Pst1 site (ctgcag->ctacag) of K362001.

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:

AhpCp2D1(Part:BBa_K1104204)
We strimmed DsbG coding out of the original sequence, making the functional sequence shorter and more efficient.

BBa_K1104204 (AhpCp2D1 promoter)Composed of AhpCp2, DsbGp, and AhpCp1. DsbG coding sequence of AhpCp1000 was trimmed out.

AhpCpD1

BBa_K1104206 (AhpCpD1 promoter)This is a bidirectional design, Composed of DsbGp(reverse) and AhpCp1(forward), and a shared TFBS for OxyR.