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

Outlines

• 1. Introduction
• 2. Experimental Design & Procedure
• 3. Results & Discussion

4.Human Practice

3

News

【 the Nanhai residents were poisoned by

• rganophosphate.

Cucumber is the culprit? 】 Nanhai had two cases of food poisoning incidents in a row. Patients were diagnosed with

• rganophosphorus

pesticide poisoning. It was suspected the poison coming from cucumber.

4

Issues with Pesticide Over Usage: Carcinogen

HELP

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

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Questionnaire

Suggested Solutions

Immerse in water Choose low toxic

• rganic pesticide

Reduce or Stop pesticide usage

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Where can you buy safe food?

A)Imported B)Domestic C)No Where D)Other A B C D

Suggested Solutions

Immerse in water Choose low toxic

• rganic pesticide

Degradate pesticide residues Reduce or Stop pesticide usage

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OpdA, a Enzyme to Degrade Organophosophorus Pesticide

( Jackson CJ, et al. Toxicology. 2004; Ely F, et al. Biochem J. 2010)

Project Introduction

OPDA

Enzyme

OmpA PEPTIDE

Experimental Goal to Achieve

Z

Z

Z

Z

Z

Z

Z

Z

Z ＜ 32 ℃

＞ 32 ℃

ultraviolet ray

32 ℃ ON/OFF

ccdB SUICIDE GENE

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Experimental Design

F1 F2 F3 F4

Experimental Procedure: PCR

F1R F1F

5’ 3’

Forward

3’ 5’

Reverse

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Experimental Procedure: Summary

Experimental Procedure: Fragment Insertion

PSB1C3 EcoRI PSTI EcoRI PSTI

Experimental Results: Inserted Segments

1 2 3 4 M PCR amplification result of F1(track 1), F2(track 2), F3(track 3) and F4(track 4)

• segments. M: DL2000

DNA marker PCR amplification result of F1+2 fragments after PCR gel extraction(track 2). M: DL2000 DNA

marker

PCR amplification result of F1+2+3+4 segments (track 2). M: DL2000 DNA

marker

PCR amplification result of F3+F4 segments (track 2). M: DL2000 DNA

marker

M

2

M

2

M

2

2000 1000 750 500 250 150

Experimental Results: Plasmid Verification

The bacterial colony photo of ligation product of pSB1C3 linear plasmid and F1+2+3+4 segments transforming into DH5α competent cells after 24 hours

Verification result of two positive plasmids after double digestion. M: λ- T14 DNA marker; track 2: positive plasmid 1 digested by PstI enzyme; track 3: positive plasmid 1 digested by EcoRI + PstI enzymes; track 4: positive plasmid 1 without digestion (negative control); track 5: positive plasmid 2 digested by PstI enzyme; track 6: positiveplasmid 2 digested by EcoRI + PstI enzymes; track 7: positive plasmid 2 without digestion (negative control)

M 2 3 4 5 6 7

7743 6223 4254 3472 2690 1882 1489

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Experimental Results: Verified by Sequencing

Sequencing results of primer VF2 Sequencing results of primer VR

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

IGEM Magazine IGEM Community IGEM Sina Weibo 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

• n synthetic biology, its ethics and safety.

Human Practice：IGEM T-shirt Theme

IGEM Special Issue on World, our School Magazine

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Wiki

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

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Acknowledgement：Special Thanks

LZU_China

Acknowledgement

Institute of Applied

• Ecology. Chinese

Academy of Sciences

Team: Shiyan_SY_China Captain：Jinyan Chu Members:Yiyang Liu, Fanglin Zhao, Kechun Zhao, Sijian Chen, Zhehao Dai, Pai Qu, Rui Zhu, Jiacong Gu Instructors: Mingkai Xu, PhD Phoebe Zhang, PhD Shuan Zhang Xiaofan Gao Xiuyu Wang, PhD

High School： (Liaoning Province Shiyan High School) Liaoning University

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