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Minesweeper Plants
Detection of landmines using transgenic Plants
Dr. SohirEl-KhodaryProfessor of Cytology & GeneticsProfessor of Molecular BiologyDr. Maher ShehataBy : Ahmed Yusuf
Supervisors:
Item 5
Different Biosensing methods used in landmine detection
Two Approaches of using Transgenic Plants for detection of landmines
•ContentsThe Points to be discussed
The problem of landmines In Egypt
Budget and Plan
Material and method
Item 1
Item 2
Item 3
Item 4
• Problem of land mines
110 million land mines in the
world
Problem of landmines in Egypt
22 Million mines !
!20% of total land mines in the
world !!
In Egypt only
5000 killed
20,000 injuredeach year
HOW TO SAFE THE WORLD?These mines must be eliminated!
DETECTION OF LANDMINES USING TRANSGENIC PLANT
OBJECTIVE
Identification of the general location of suspected area
Accurately defining the outer perimeters of the landmine contaminated areas
Actual Clearance of the area Identified in the previous step
Demining Process
1
23
Methods of Demining
PlantsBacteria
BeesRats
Dogs
Demining using Biosensors
transgenic Plants as Minesweeper
Promoter FusionSynthetic Receptor
There are two strategies
PROMOTER FUSIONThe First Strategy
(A) Promoter fusion Strategy
Gene X
Expressed regionPromoter
Red pigment in leaf
(A) Promoter fusion Strategy
Gene Y
Nitrate reductase
TNT
(A) Promoter fusion Strategy
Gene Y
NR
TNT
Red pigment in
leaf
Gene XGene Z
Aresa® Biodetection Technology(A )Promoter Fusion
disadvantage
Naturally found in
soil
• Arabidopsis microarray reveal genes activated by these four volatiles:2,4-DNT, 2,4,6-TNT, 2-Amino DNT and 4-Amino-DNT
• The Promoters of these genes could be used instead of NR genes
• Unfortunately These four genes are also induced by naturally occurring stress and natural plant hormones
Refinement of This TechnologyO2N
NO2
NO2 O2N
NO2
NH2
O2N
NH2
NO2 O2N
NO2
To overcome this problem..
Logic gate : AND
e.g. Selected promoters could each be used to direct transcription of a different necessary component of a reporter system, such as a series of genes in a pigment biosynthetic pathway.
SYNTHETIC RECEPTORThe second Strategy
(B )Synthetic receptor Strategy
Synthetic PBPSynthetic ST
circuitDe-greening
response
Animation
(B )Synthetic receptor Strategy
BudgetTotal(LE) Details Item
36,00096,00036,000
1 person x LE 1500 x 24 months4 person x LE 1000 x 24 months3 person x LE 500 x 24 months
Staff Salaries Project PIProject Co-IResearchers
30,0004,0002,0008,00040,0002,00020,0002,00040,000
120,000
PCRDNA electrophoresisDigital pH meterGlassdistillator(water apparatus)Gel documentation systemHot plate with magnetic stirrerMicrocentrifugeVotexUV spectrophotometerGene gun
Equipments
50,000
DNA extraction kits, plasmids, DNA polymerases, primers, enzymes, agarose, Biolistic gun kits.
Chemicals
10,000 Sampling equipments Field consumables50,000 Field requirements Field trips
30,000 Local, Regional & International Conferences& Workshops
5,000 Papers and repots Publication
10,000 Others
LE 591,000
Total Budget
Months Tasks
I-Collection of plant material and plasmid constructs
II-Production of transgenic plants & statistical analysis
III- Field work
Plan
1-6 7-12 13-18 19-24
• Collection of samples.• Microarray of plants treated with explosives.• Promoter analysis of genes induced by explosives.• Promoter fusion with reporter gene.
Materials and Methods
• Optimization and adaptation of the transformation protocol (Host-mediated by Agrobacterium or Non-host mediated by Biolistic gun)
• Gene bombardment to the selected tissues using Biolistic Gun
• Selection of successfully transformed tissues• Molecular characterization of transformants • Regeneration of transgenic plants expressing new genes
Materials and Methods
• Growing of transgenic plants and test the stability of gene in progenies, and then test in field
• 1st challenge experiment (Growth room evaluation)• 2nd challenge experiment (Greenhouse evaluation)• 3rd challenge experiment (Field evaluation)• Multi-location and multi- year evaluation of the progeny• Introducing of transgenic seeds in minefield using plane or helicopter
Materials and Methods
ACKNOWLEDGEMENT
THANK YOU
