A riboswitch controlled location- based networking framework - - PowerPoint PPT Presentation

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A riboswitch controlled location- based networking framework - - PowerPoint PPT Presentation

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A riboswitch controlled location- based networking framework Bacterial chemotaxis Chemotaxis: chez-mediated flagellar rotation o vervi erview: ew: de design ign Simulation ation Ch Characteri acterizatio zation Outr treach ach

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A riboswitch controlled location- based networking framework

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Bacterial chemotaxis

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Chemotaxis: chez-mediated flagellar rotation

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  • vervi

erview: ew: de design ign

Simulation ation Ch Characteri acterizatio zation Outr treach ach Summar mary y & ac achievements vements Acknowle

  • wledge

dgeme ments nts

TH AN K YO U
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SLIDE 9

Design Construction

CheZ-reporter

Function

START END

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SLIDE 10

START

Analyte A Analyte B

Analyte A gradient Analyte B gradient

Project Design & Construction

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Analyte A Analyte B

Send & Receive motility module

Riboswitch B

CheZ-mRFP CheZ-Venus

Riboswitch A

lox66 lox71

Pon

B

CheZ-Venus

towards A

A

START

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SLIDE 12

Riboswitch B

CheZ-mRFP CheZ-Venus

Riboswitch A

lox66 lox71

Pon

B

CheZ-Venus

towards A

A

Cre-activating toggle switch

START Analyte A Analyte B

PX PY

X Cre

Y X

Y

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SLIDE 13

Riboswitch B

CheZ-mRFP CheZ-Venus

Riboswitch A

B

Analyte A Analyte B

PX PY

X Cre

Y X

Y

Cre

lox66

Pon

lox71

CheZ-mRFP

towards B

A

START

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Atra razine zine rib iboswitch switch

Atraz azin ine e ribos

  • switc

witch

(Sinha ha and and Gallivan livan, NCB NCB, , 2010) 10)

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8.1 Theophy phylli line ne Ribos

  • switch

witch

(Topp pp and and Galliva ivan, , JACS, , 2007) )

Theophyll hylline ne ribosw switch itch 1

Riboswitches

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12.1 1 Theophy phylline lline ribos

  • switc

witch

(Lynch ch and Gallivan ivan, , NAR, 2009)

Theophyll hylline ne ribosw switch itch 2

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CheZ

WITS_CSIR thRS-CheZ-reporter

Venus mRFP

CDS “x”

theophylline riboswitch BBa_K249028 Lethbridge

11nt insert

theophylline riboswitch BBa_K411001 Taipei

6nt scar

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Overvie rview sim

imulation ulation

Ch Characteri acterizatio zation Outr treach ach Summar mary y & ac achievements vements Acknowle

  • wledge

dgeme ments nts

TH AN K YO U
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Overvie rview: w: design gn Simulation ation

cha haract racterizat erization ion

Outr treach ach Summar mary y & ac achievements vements Acknowle

  • wledge

dgeme ments nts

TH AN K YO U
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Empty vector Riboswitch 1

Venus RS1

BBa_K537009 Riboswitch 2

Venus RS2

BBa_K537010

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Riboswitch 1 chez venus

8-fold activation

CheZ Venus RS1

BBa_K537011

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Riboswitch 2 chez venus

46-fold activation

CheZ Venus RS2

K537012

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SLIDE 24

CheZ Venus RS1

Riboswitch 1 CheZ

CheZ Venus RS2

Riboswitch 2 CheZ

BBa_K537011 BBa_K537012

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BBa_K537012 BBa_K537011 5-fold difference

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Riboswitch 1 Riboswitch 2

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Overview rview Simulation ation Ch Characteri acterizatio zation Outreach

utreach

Summar mary y & ac achievements vements Acknowle

  • wledge

dgeme ments nts

Th ank Yo u
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Wor

  • rksho

kshops ps at Sci-bon bono

  • disc

scove very y centre ntre: “national science week” 2-6 6 Au Augus ust 2011 11

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+

‘Superhero’ bacteria!

Instruction

Problem Solution

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Overview rview Simulation ation Ch Characteri acterizatio zation Outr treach ach Summar

ary y & a & achie ievem vemen ents

Acknowle

  • wledge

dgeme ments nts

TH AN K YO U
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Submitted 11 new parts to the registry Fully characterized two theophylline riboswitches Collaborated with ICL on modelling and assays Introduced synthetic biology to the broader public

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A finalist at the European Jamboree Improved the aesthetics and content of wiki Improved experiments & microscopy

Had lots of fun!

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Overview rview Si Simulation lation Ch Characteri acterizatio zation Outr treach ach Summar mary y & ac achievements vements Ackn

knowledgement

  • wledgements

THANK YOU

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SLIDE 35
  • Prof Kramer
  • Prof Patrick Arbuthnot
  • Prof Theresa Coetzer
  • Dr Sylvia Fanucchi
  • Prof Heini Dirr
  • David Balchin
  • Robyn Brackin
  • The Synthetic Biology Research Unit at the

CSIR

  • Dr Sonja Lauterbach
  • Kubendran Naidoo
  • Dr Toru Nakayashiki and Prof Hirotada Mori
  • Dr Joy Sinha and Prof Justin Gallivan

Supervisors

  • Karl Rumbold
  • Marco Weinberg
  • Musa Mhlanga
  • Raymond Sparrow

Advisors

  • Michelle Robinson
  • Laura Millroy
  • Youtaro Shibayama
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SLIDE 36