SLIDE 1
S R I E N S
ynthetic anger ntelligent ematode .coli
Agianst
S I R E N S ntelligent .coli ynthetic ematode anger Agianst - - PowerPoint PPT Presentation
S I R E N S ntelligent .coli ynthetic ematode anger Agianst - - PowerPoint PPT Presentation
S I R E N S ntelligent .coli ynthetic ematode anger Agianst 1 Economic damage Environment hazard lack of weariness background Meloisogyne spp. [1] Meloisogyne incoginta [2] Heterodera spp. [1] Bursaphelenchus xylophilus [1] [1]
SLIDE 2
SLIDE 3
background
1
Economic damage
lack of weariness Environment hazard
Meloisogyne spp.[1] Bursaphelenchus xylophilus[1] Heterodera spp.[1] Meloisogyne incoginta [2]
[1] wikipidia [2] Beijing Agricultural database SLIDE 4
SLIDE 5
3rd announcement of China’s State Forestry Administration (SAF) in 2015
HALF
CHINA!
- f
background
1
Economic damage
lack of weariness Environment hazard
SLIDE 6
Common Nematicides
Nematicides Targets Disadvantages Ethylene dibromide Nematodes in the soil Other small pests High toxicity. Do harm to the soil, the environment even the health. The using period is limited. It is probably do harm to the crops. Preventive nematicides. The operation is very complex. Fenamiphos Apelenchoides fragariae Most of the nematodes Have toxicity. The underground water will be polluted. Abamectin Meloidogyne; Pratylenchus; Radopholus; Tylenchulus; Heterodera glycines Expensive Biological agent DiT Meloidogyne; Heterodera; Pratyleachus spp.; Trichodorus spp.; Belonolaimus spp.
background
1
Economic damage lack of weariness
Environment hazard
SLIDE 7
Villagers only care about visible pests instead of invisible nematodes Officials and local pesticide retailer knew little about nematodes
Economic damage
Lack of awareness
Environment hazard
Investigation in Hebei Province background
1
SLIDE 8
Sample analysis - Soil nematode screening
Economic damage
Lack of awareness
Environment hazard
Investigation in Hebei Province background
1
SLIDE 9
Communication With Other Teams The abuse of pesticide have brought about severe agricultural diseases all over the world! background
1
Economic damage
Lack of awareness
Environment hazard
SLIDE 10
An interview with Dr. Yan Houfu (Beijing Normal University) Lack of bills on bio-safety, Lack of protection of farmers’ rights! background
1
Economic damage
Lack of awareness
Environment hazard
SLIDE 11
We aim to solve these problems ! How to solve these problems ?
?
Economic damage of nematodes Environmental hazard The abuse of pesticide have brought about severe agricultural diseases all over the world! Lack of bills on bio-safety Officials and local pesticide retailer knew little about nematodes Villagers only care about visible pests instead of invisible nematodes Lack of protection of farmers’ rights
SLIDE 12
?
SLIDE 13
SLIDE 14
The !Project
SLIDE 15
Bait & Killer System
Overview
Photoinduced System BDT System
Photo-induced System Bidirectional Transcription System Bait & Killer System Suicide Project
2
Suicide
SLIDE 16
The construction result of light regulation system Cph8
Project
2
Bait & Killer System Overview
Photoinduced System
BDT System Suicide
SLIDE 17
Detection experiment result Biobrick 1 & 2 Biobrick 1 Biobrick 2
Project
2
Bait & Killer System Overview Photoinduced System
BDT System
Suicide
SLIDE 18
Project
2
SLIDE 19
terpenoid attracts nematodes such as Tylenchulus semipenetrans
d-limonene l-limonene
Limonene Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 20
Engineered pathway for l-limonene biosynthesis in E. coli
Biosysthesis Pathway
Reference : Du et al. Bioresources and Bioprocessing,2014,1:10.Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 21
Vector : pGEX-4T-1 plasmid Chassis : E. coli BL21 (DE3)
Promoter RBS GPPS LS
BBa_J23100 Strong constitutive promoter BBa_B0032 GPPS gene : Abies grandis (GenBank : AF513112) d-LS gene : Citrus unshiu (GenBank : AB110636.1) l-LS gene : Mentha spicata (GenBank : L13459)
Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 22
Limonene Production
SDS-PAGE of l-/d- limonene synthase and GPPS
LS : 89kDa GPPS: 49kDa
d-limonene GC-MS
Agarose gel electrophoresis
Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 23
Proportion of nematodes' distribution The distribution of C. elegans
Limonene attracts C.elegans significantly Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 24
Consulting with Prof. Liu Xingzhong (PhD, CAS) Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 25
Chassis : E. coli BL21(DE3)
RBS bace16 / rMpL Promoter
BBa_K206000 pBAD promoter BBa_B0032 bace16 gene : Bacillus nematocida (GenBank : AY708655) rMpL gene : Macrolepiota procera (GenBank : HQ449739)
Promoter
BBa_J23100 Constitutive promoter
Vector : pSB1C3 plasmid Chassis : E. coli BW25113
Toxins: Bace16 & rMpL
Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 26
rMpL successfully expressed Project
2
SDS-PAGE of rMpL
Lane 1, supernatant of pSB1C3-BL21(DE3) with constitutive promoter; Lane 2, supernatant of pSB1C3-BW25113 with pBAD promoter after 0uM L-Ara induction; Lane 3, supernatant of pSB1C3.
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 27
rMpL stops the C.elegans growth
rMpL PSB1C3 empty plasmid
Project
2
Bait & Killer System
Overview Photoinduced System BDT System Suicide
SLIDE 28
AHL LuxR LuxR-AHL LuxI
Project
2
Bait & Killer System Overview Photoinduced System BDT System
Suicide
SLIDE 29
Device&Modeling
SLIDE 30
Device
Modeling
slide glasses
- uter shell
red LED light separating funnel test tube
CO2 Generating Chamber E.coli Culturing Chamber Light Controlling Chamber Cuboid Outer Shell
Device&Modeling
3
SLIDE 31
http://2015.igem.org/wiki/images/0/00/BNU-MODELING-DEVICE-VIDEO.mp4
Device&Modeling
3
Device
Modeling
SLIDE 32
dichotomy
The longest distance between nematodes and the equipment Distance between two devices Diffusion coefficient of limonene gas diffusion model the concentration
- f every device
Nematodes’ movement velocity, attraction time
L = V × T
AB = 2 √ 2EF
C0 2 (1 − erf x 2 √ Dt )
= C0 2 (1 −
18X
i=0(−1)i
x2i+1 i!(2i + 1)(2 √ Dt)2i+1 )
[1] Limm W, Begley T H, Lickly T, et al. Diffusion of limonene inpolyethylene[J]. Food additives and contaminants, 2006, 23(7): 738-746.
∂C(t, x) ∂t = DA ∂2C(t, x) ∂x2
Fick’s Law:
V: nematodes’ crawl speed T: the half time of which nematodes grow from eggs to larvae
To Find The Best Position For The Device To Be Placed In Farmland
simulation cellular automaton Device
Modeling
Device&Modeling
3
SLIDE 33
Database
extension
4
Public engagement
Voluntary education in GuiZhou Province Synthetic biology Plant diseases and insect pests SIRENS
SLIDE 34
Database
Public engagement
extension
4
Open day of BNU-CHINA Synthetic biology iGEM competition SIRENS Interviewed by BNU TV station Our innovative idea How iGEM impact us Our achievements
SLIDE 35
extension
4
Database
Public engagement
SLIDE 36
Wet lab Modeling Policy & Practice Wiki Art Team
5
SLIDE 37
Ackonwledgement
6
SLIDE 38
Thanks！
SLIDE 39