Euk.cement - 2015.igem.org2015.igem.org/files/presentation/HUST-China.pdfenvironmentally safe...
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Euk.cement
Transcript of Euk.cement - 2015.igem.org2015.igem.org/files/presentation/HUST-China.pdfenvironmentally safe...
Euk.cement
Who am I?
A Journey
Desert in Northwest China
A Journey
Hangzhou, China
A Journey
Dead coral, Sanya, China
Incompact sand substrates!
This is the key issue for those problems…
Consolidate!
Traditional methods Biological methods
Time for show!Promotion of micro-consolidation tech
environmentally safe microorganism
live in marine environment
hydrophobic substrate
ability to grow fast
high-density fermentation
express complex recombinant proteins
Yarrowia Lypolitica
LIP2 prepro Mcfp-3
BD-CRY2 AD-CIB1
UAS
PTEF PADH1
Pgal1 Panb1
ROX1 LIP2 prepro Si-tag YLcwp3
Panb1
Genetic circuits
• 1. darkness induction system
• 2. silica binding system
• 3. flocculating system
GalBDCRY2
Gal4 UAS
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GalBDCRY2
Gal4 UAS
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CIB GalAD
CIBGalAD
light
dark
1. Darkness induction system-- Light control system based on Y2H
BD-CRY2 AD-CIB1
UAS ROX1
pTEFpADH1
pGal1 pAnb1
LIP2 prepro Si-tag YLCwp3
LIP2 prepro Mcfp-3
pAnb1
XPR2 Mcfp-3
CRY2-BD CIB1-AD
UAS ROX1
pTEFpADH1
pGal1 pAnb1
XPR2 Si-tag Y1CwpC
pAnb1
1. Dark induction system
Panb1
LIP2 prepro Si-tag YLcwp3
2. Silica binding system
Si-tag
cell
Silicon surface
2. Silica binding system
LIP2 prepro Mcfp-3
pAnb1
3. Flocculating system
Modeling & experiments
Modeling & experiment
Ms. Experiment
Mr. Modeling
Light control systemsmodeling
ModelingLight control systems
Modeling
Surface display & secret system
Fusion expression of 6xHis tag
Experiment • Immunofluorence
• Surface display
Silica binding testExperiment • Glass surface binding
• Three different intensity
Flocculating of secreted Mcfp-3
Experiment • Successful secretion
• Successful flocculating
Consolidation test
Experiment • Mimetic seabed• Successful cementation
Prof. J.J.Zheng Huazhong University of Sci & Tech
(HUST), Wuhan, China
Expert in civil engineering
Experiment
Offer harbors to marine organisms
Modeling
Simulation of darkness induction system
Modeling
Diffusion models
• Fast diffusion• Semi-circle pattern
ModelingDesign for diffusion
Safety
Implementation method revisited
Safety
Container
“Fence”
Prof. Weiding Wang
Marine Fisheries Research Institute,
Zhejiang, ChinaExpert in aquafarm and artificial reefs
Human Practice
Investigation in Zhoushan Islands, China
Human Practice
Human Practice
Exchanges and conferences in
• UC Berkeley @CA
• PKU @Beijing
• NCTU @Taiwan
• WHU @Wuhan
Parts submittedPart Number Description Abbrevi
ation
BBa_K1592005 GalBD-CRY2 Fusion forYeast-Two-Hybrid
BD-CRY2
BBa_K1592006 GalAD-CIB1 Fusion forYeast-Two-Hybrid
AD-CIB1
BBa_K1592015 photoreceptorcryptochrome 2
CRY2
BBa_K1592016 a basic helix-loop-helixprotein
CIB1
Light control parts
Part Number Description Abbreviation
BBa_K1592007 LIP prepro + E. coli ribosomalprotein L2 (1-60) + YLcwp3 Fusion
Si-tag1-his
BBa_K1592008 LIP prepro + E. coli ribosomalprotein L2 (61-202) + YLcwp3 Fusion
Si-tag2-his
BBa_K1592009 LIP2 prepro + E. coli ribosomalprotein L2 (203-273) + YLcwp3 Fusion
Si-tag3-his
BBa_K1592010 LIP2 prepro + E. coli ribosomal proteinL2 (1-202)+ YLcwp3 Fusion
ST12
BBa_K1592011 LIP prepro + E. coli ribosomal protein L2(61-273) + YLcwp3 Fusion
ST23
BBa_K1592012 LIP prepro + E. coli ribosomal protein L2(1-60,203-273) + YLcwp3 Fusion
ST13
BBa_K1592013 LIP prepro + E. coli ribosomal protein L2(1-60,GS linker,202-273) + YLcwp3
ST1L3-his
BBa_K1592014 LIP prepro + E. coli ribosomalprotein L2 (1-273) + YLcwp3
ST123
Silica-tag protein
PartNumber
Description Abbreviation
BBa_K1592001
Mytilus californianusfoot protein 3(Mcfp3)variant 3
Mcfp3
BBa_K1592003
Mcfp3 with LIP2prepro
LIP-Mcfp
BBa_K1592017
Mcfp3 with XPR2pre
XPR2-Mcfp
Flocculent protein
Part Number Description Abbreviation
BBa_K1592020 Ptrp mutant1 Ptrp1
BBa_K1592021 Ptrp mutant2 Ptrp2
BBa_K1592022 Ptrp mutant3 Ptrp3
Part Number DescriptionBBa_K1592018 Pgal1+rox1+cyc1_terminatorBBa_K1592019 Panb1+XPR2 pre-Mcfp3BBa_K1592023 Ptrp mutant1+RBS+GFPBBa_K1592024 Ptrp mutant2+RBS+GFPBBa_K1592025 Ptrp mutant3+RBS+GFP
PartNumber
Description Abbreviation
BBa_K1592000
LIP2 prepro(signal peptide) LIP2prepro
BBa_K1592002
Yarrowia lipolytica cell wallprotein 3
YLcwp3
Secrete and surface display
OthersPart Number Description Abbreviation
BBa_K1592004 promoter hp4d Php4d
Parts improved
Composite parts
Parts submitted
Achievements
Register, complete the judging form, wiki, poster and presentation, new BioBrick
Validated new BioBrick
Documented characterization of new parts and submit them
Abundant and effective human practice which is tightly related to our project
Cooperated with WHU & HZAU
Improved and further documented some previous parts
advisors: Professor Y.J. Ya,Professor K.Ning,Dr Y. Zhan,Dr Q. Gewetlab: S.Y. Tang, L. Yin, Z.Y. Cheng, W.Xi, W.J Liu,J.J. Xu, S.Y. Xiang, Z.Zeng, Y.X. Zheng and G.Z. Wu.Wiki framework:Y. XuArt and design:Z.X. Yangvideo:Y.X. Zheng J.Z. YangHP:J.T. Liu, S.Y. Tang,G.Z. Wu,Professor X.Q. Zhao and Professor Jens Nielson providing Saccharomyces cerevisiae s288c.Thanks Drs and masters for their direct guide in experiment.Thanks Dr X.H. Gui, M Yulin Kuang Dr G.L. Wang, Dr W. Liu, Dr D.J. Pan R.Y. Zhang
Acknowledgements
Team
Thank you!
