Pesticide Residue Killer iGEM Jamboree Liaoning Province Shiyan - PowerPoint PPT Presentation

Pesticide Residue Killer ——iGEM Jamboree Liaoning Province Shiyan High School Shiyan_SY_China 9/25/2015

1. Introduction 2. Experimental Design & Procedure Outlines 3. Results & Discussion 4.Human Practice

News 【 the Nanhai residents were poisoned by organophosphate. Cucumber is the culprit? 】 Nanhai had two cases of food poisoning incidents in a row. Patients were diagnosed with organophosphorus pesticide poisoning. It was suspected the poison coming from cucumber. 3

Issues with Pesticide Over Usage: Carcinogen HELP 4

The challenge: Rising population but Limited farm land Pesticide is necessary to provide sufficient food supply 6

Questionnaire 7

Suggested Solutions Choose low toxic Reduce or Stop Immerse in water organic pesticide pesticide usage

Where can you buy safe food? A)Imported B)Domestic C)No Where D)Other A B C D 9

Suggested Solutions Choose low toxic Reduce or Stop Immerse in water organic pesticide pesticide usage Degradate pesticide residues

Project Introduction OPDA OmpA PEPTIDE Enzyme OpdA, a Enzyme to Degrade Organophosophorus Pesticide ( Jackson CJ, et al. Toxicology. 2004; Ely F, et al. Biochem J. 2010) 11

Experimental Goal to Achieve 32 ℃ ON / OFF Z Z Z Z ＜ Z Z ultraviolet ray 32 ＞ ℃ 32 ℃ Z Z ccdB Z SUICIDE GENE

Experimental Design F1 F2 F3 F4 13

Experimental Procedure: PCR F 1 R 3’ Forward 5’ Reverse 5’ 3’ F 1 F

Experimental Procedure: Summary 15

Experimental Procedure: Fragment Insertion EcoRI PSTI EcoRI PSTI PSB1C3

Experimental Results: Inserted Segments 2000 1000 750 500 250 150 M M M 1 2 3 4 M 2 2 2 PCR amplification PCR amplification PCR amplification PCR amplification result of F1(track 1), result of F1+2 result of F3+F4 result of F1+2+3+4 F2(track 2), F3(track fragments after PCR segments (track 2). segments (track 2). 3) and F4(track 4) gel extraction(track M: DL2000 DNA M: DL2000 DNA segments. M: DL2000 2). M: DL2000 DNA marker marker DNA marker marker

Experimental Results: Plasmid Verification 7743 6223 4254 3472 2690 1882 1489 M 2 3 4 5 6 7 The bacterial colony photo of Verification result of two positive plasmids after double digestion. M: λ - T14 DNA marker; track 2: positive plasmid 1 digested by PstI enzyme; ligation product of pSB1C3 track 3: positive plasmid 1 digested by EcoRI + PstI enzymes; track 4: linear plasmid and F1+2+3+4 positive plasmid 1 without digestion (negative control); track 5: positive segments transforming into plasmid 2 digested by PstI enzyme; track 6: positiveplasmid 2 digested by DH5α competent cells after EcoRI + PstI enzymes; track 7: positive plasmid 2 without digestion 24 hours (negative control)

Experimental Results: Verified by Sequencing Sequencing results of primer VF2 Sequencing results of primer VR 19

Conclusion & Discussion Conclusion: we had successfully constructed and submitted a plasmid that expressing temperature controlled OpdA protein in vitro and the expression can be shut down by UV induced suicide gene. Discussion: Further functional tests need to be done. • Expression dynamic curve of OpdA Secretion of OpdA • • Activation curve of suicide gene

Human Practices I GEM Sina Weibo IGEM Magazine IGEM Community IGEM Video Promotion

Human Practice: Online Survey on Pesticide Issues Yes No Don’t know

Human Practice: Video Promotion on Project The video about our project which can be found on Yuku website http://www.tudou.com/programs/view/OznKAMRUQac/

Teaching Our Peers about Synthetic Biology On July 23rd, we held a lecture on iGEM and synthetic biology in the International Department of Liaoning Province Shiyan High School. In the lecture, we introduced the historical courses of iGEM, and helped students have a preliminary understanding on what is synthetic biology, and enumerated the application of biology in our life. The students had a heated discussion about their own ideas on synthetic biology, its ethics and safety.

Human Practice ： IGEM T-shirt Theme

IGEM Special Issue on World , our School Magazine

Wiki http://2015.igem.org/Team:Shiyan_SY_China 27

Acknowledgement ： Special Thanks LZU_China 28

Acknowledgement High School ： (Liaoning Province Institute of Applied Ecology. Chinese Shiyan High School) Academy of Sciences Team: Shiyan_SY_China Instructors: Captain ： Jinyan Chu Mingkai Xu, PhD Members :Yiyang Liu, Phoebe Zhang, PhD Fanglin Zhao, Kechun Shuan Zhang Zhao, Sijian Chen, Xiaofan Gao Zhehao Dai, Pai Qu, Rui Xiuyu Wang, PhD Liaoning Zhu, Jiacong Gu University

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