Contents Team information Who are we? Where are we from? - - PowerPoint PPT Presentation

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Contents Team information Who are we? Where are we from? - - PowerPoint PPT Presentation

Mar 13, 2023 368 likes •866 views

Contents Team information Who are we? Where are we from? The Idea The issue The solution Application in society The machine Components of the machine How the machine works Further

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SLIDE 1
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SLIDE 2

Contents

  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 3
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 4

Team Information

Breegini Culas Dr Leonard Pattenden (The Mad Scientist) Muriel Aldunate Candice Raeburn Candice Raeburn Sebastian Ramov (The Bully) Michelle Chayeb (The Small One) Lyna Thach Danielle Kamato (The Shopaholic) Jasmine Panthaki Flora Yuen

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SLIDE 5
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 6
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SLIDE 7

Where we come from?

www.cityofmelbourne.com.au

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SLIDE 8
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 9

The Issue

Too much electricity use is a global concern, causes fossil fuel to be burnt

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SLIDE 10
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 11

The Solution

Electricity Fossil Fuels Green House Gas Emission Global Warming Our Machine Biological Lighting System

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SLIDE 12
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 13

Applications - Manufacturing

House Hold lighting Commercial Lighting Street Lighting Christmas Tree Lights

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SLIDE 14
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Results
  • Making a part
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 15

Components of Our Machine

  • Fluorescent Proteins
  • Cell Free Chassis
  • E. Coli Cell Free System
  • Wheat Germ Cell Free System
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SLIDE 16
  • Fluorescent Proteins
  • Cell Free Chassis
  • E. Coli Cell Free System
  • Wheat Germ Cell Free System
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SLIDE 17

Fluorescent Proteins

YFP BBa_E0030 GFP Cherry BBa_1712028 VIC_Green BBa_E0040 BFP Blueberry

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SLIDE 18

Structure Of Fluorescing Protein

α helix β-strands

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SLIDE 19

How Do They work?

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SLIDE 20 Tyr66 Ser65
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SLIDE 21
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SLIDE 22
  • Fluorescent Proteins
  • Cell Free Chassis
  • E.Coli Cell Free System
  • Wheat Germ Cell Free System
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SLIDE 23

Centrifuged supernatant

CELL FREE SYSTEMS

  • What Is a Cell Free System?

S

  • n

i c a t i

  • n

Cleared Lysate

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SLIDE 24
  • How Does A Cell Free System

Work?

  • reactions that occur in a cell take

place in a cell free system with

  • ut the complex interactions found

in the whole cell

http://genome.imim.es/

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SLIDE 25
  • Types of Cell Free Systems
  • Wheat Germ; safer alternative - non toxic

www.hondaireland.ie

Endogenous mRNA= >protein synthesis Manufacturing, sustainability

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SLIDE 26
  • Why A Cell Free System?

From: Collections of Henry Ford P189.16783

Potential ordered and sequential control

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SLIDE 27
  • not optimised - scale limited
  • Synthetic biology; cloning is defined

and standardised

  • Can optimise the chassis and move to

scale

  • control the machine
  • components and toxins
  • Non propagating and non replicating
  • E.coli ideal for coupled reactions, simple
  • ethicals

less complication at control of initiation means better synthesis

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SLIDE 28

Resuspended pallets S30 A buffer Centrifuged 2000rpm Resuspended pallets S30 B buffer Repeated 3 times Sonicated Centrifuged 20000 x g Supernatant was E.coli Extract

S30 Buffer: Potassium Acetate, Magnesium Sulphate, MQ water pH of 7.8 S30 A contained 2mercaptoethanol

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SLIDE 29
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 30

How does the Machine Work?

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SLIDE 31
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 32

Where to next?

  • Switching on and off
  • Resonance Energy transfer
  • Fluorescence Resonance Energy Transfer

(FRET)

  • Bioluminescent Resonance Energy Transfer

(BRET)

  • Scale up
  • Dialysis
  • Continuous Exchange Cell Free (CEFC)
  • Continuous Flow Cell Free (CFCF)
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SLIDE 33

Resonance Energy Transfer

  • Distant dependant

energy transfer

  • FRET- external

illumination

  • BRET- luciferase

(oxidative enzyme) Energy Transfer with the emission

  • f a photon
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SLIDE 34

Dialysis

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SLIDE 35
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 36

Making the Part

  • iGEM fluorescence protein

Biobricks in many colours but no blue

  • Different strategy of cloning:

Quikchange Mutagenesis: point Mutations

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SLIDE 37

Quikchange Mutagenesis

Mutant Strand Synthesis: Mutant Strand Synthesis:

  • Denature DNA template
  • Anneal mutagenic primers
  • Extend primers

Dpn Dpn I Digestion of Template I Digestion of Template

  • Digest parental methylated

DNA Transformation: Transformation:

  • Transform mutated

plasmid

7 mins 68 1 3 1 min 68 18 2 50 sec 60 50 sec 95 1 min 95 1 1 time Temp Cyc Seg

Plasmid DNA is methylated

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SLIDE 38
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 39

Results

  • Expressing BFP,

GFP,YFP and cherry

  • Quick Change Failed
  • No visible

fluorescence in machine

  • Fluorescence still

visible after 8 weeks

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SLIDE 40
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next Year
  • Acknowledgements
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SLIDE 41

Problems!!

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SLIDE 42
  • Experimental Problems
  • Leaving PCR reaction on the sink
  • ver the weekend
  • Mix up in what reagents were

added to the reactions

  • Pipetting
  • clumsiness
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SLIDE 43
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 44

Next year…

  • First year in iGEM
  • Learnt how things are run and

how things should be done

  • That means we will be back

next year with our aims set at gold

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SLIDE 45
  • Team information
  • Who are we?
  • Where are we from?
  • The Idea
  • The issue
  • The solution
  • Application in society
  • The machine
  • Components of the machine
  • How the machine works
  • Further ideas?
  • Making a part
  • Results
  • Problems and limitations
  • Next year
  • Acknowledgements
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SLIDE 46

Acknowledgements

  • We would like to thank the following for their help and

contributions to our project

  • Friends, family and teachers who provided moral support and

guidance to the teams' Undergraduate students

  • Rushdi Mussa (friend): was a great help in designing the wiki

page and putting it together.

  • Mr Kieran Wilson: Web Content Coordinator for the Science,

Engineering & Health Office (RMIT University)

  • Mr Robin Dick: Manager, Benefit Attraction. Alumni and

Development (RMIT University)

  • Bennetts Lane Jazz, Farm pro, Journal Cafe and Journal

Canteen for their donations

  • Prof David Adams (Director Health Innovations Research

Institute

  • Prof Peter Coloe PVC School of Science, Engineering and

Health

  • Prof Margaret Gardiner Vice Chancellor RMIT University
  • School of Medical Science Finance Dept.
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SLIDE 47

Sponsors

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SLIDE 48
  • THANK YOU

iGEM !!