FLUORESCENT REPORTER CHARACTERIZATION Judith Eeckman, Adam Grose, - - PowerPoint PPT Presentation

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FLUORESCENT REPORTER CHARACTERIZATION Judith Eeckman, Adam Grose, - - PowerPoint PPT Presentation

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FLUORESCENT REPORTER CHARACTERIZATION Judith Eeckman, Adam Grose, Miranda Hagen, Brittany Harwell, Swetha Pasala, Meghan Savage, Hayley Schaefer, Loran Steinberger, Mark Stewart, Daniel Thorpe Laura Adam, Dr. David Ball, Dr. Martha Eborall,

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SLIDE 1

FLUORESCENT REPORTER CHARACTERIZATION

Judith Eeckman, Adam Grose, Miranda Hagen, Brittany Harwell, Swetha Pasala, Meghan Savage, Hayley Schaefer, Loran Steinberger, Mark Stewart, Daniel Thorpe Laura Adam, Dr. David Ball, Dr. Martha Eborall, Julie Marchand, Dr. Jean Peccoud

2011

Sunday, October 9, 2011

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SLIDE 2

2011 Introduction

TetR λ cl LacI

Sunday, October 9, 2011

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SLIDE 3

2011 Introduction

TetR λ cl LacI

Sunday, October 9, 2011

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SLIDE 4

2011 Introduction

TetR λ cl LacI

Sunday, October 9, 2011

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SLIDE 5

2011 Introduction

TetR λ cl LacI

Sunday, October 9, 2011

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SLIDE 6

2011 Introduction

TetR λ cl LacI

Sunday, October 9, 2011

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SLIDE 7

2011 Introduction

Elowitz and Leibler 2000

TetR λ cl LacI

Sunday, October 9, 2011

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SLIDE 8

2011 Objectives

  • 1. Characterize fluorescent reporters
  • 2. Look for context-dependence
  • 3. Explore the problems of parts characterization

Sunday, October 9, 2011

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SLIDE 9

2011 Current methods

In vivo vs. In vitro

http://www.mbl.edu/news/features/shimomura.html Credit: Tom Kleindenst

Sunday, October 9, 2011

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SLIDE 10

2011 Outline

Sunday, October 9, 2011

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SLIDE 11

2011 Outline

Design

Sunday, October 9, 2011

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SLIDE 12

2011 Outline

Design

Sunday, October 9, 2011

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SLIDE 13

2011 Outline

Design Fabrication

Sunday, October 9, 2011

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SLIDE 14

2011 Outline

Design Fabrication

Sunday, October 9, 2011

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SLIDE 15

2011 Outline

Design Fabrication

Characterization

Sunday, October 9, 2011

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SLIDE 16

2011 Outline

Design Fabrication

Characterization

Sunday, October 9, 2011

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SLIDE 17

2011 Outline

Design Fabrication

Characterization

Analysis

Sunday, October 9, 2011

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SLIDE 18

2011 Outline

Design Fabrication

Characterization

Analysis

Sunday, October 9, 2011

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SLIDE 19

2011 Outline

Design Fabrication

Characterization

Analysis

Sunday, October 9, 2011

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SLIDE 20

2011 Outline

Design Fabrication

Characterization

Analysis

Sunday, October 9, 2011

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SLIDE 21

Design Fabrication Analysis

2011

Characterization

Roadmap

Name Tag Organism Design Cloning Screening Sequencing Imaging Data Analysis

AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine None

  • S. cerevisiae

None

  • S. cerevisiae

None

  • S. cerevisiae

None

  • S. cerevisiae

None

  • S. cerevisiae

None

  • S. cerevisiae

None

  • E. coli

None

  • E. coli

None

  • E. coli

None

  • E. coli

None

  • E. coli

PEST

  • S. cerevisiae

PEST

  • S. cerevisiae

PEST

  • S. cerevisiae

PEST

  • S. cerevisiae

PEST

  • S. cerevisiae

PEST

  • S. cerevisiae

Ubi-Lys

  • S. cerevisiae

Ubi-Lys

  • S. cerevisiae

Ubi-Lys

  • S. cerevisiae

Ubi-Lys

  • S. cerevisiae

Ubi-Lys

  • S. cerevisiae

Ubi-Lys

  • S. cerevisiae

LVA

  • E. coli

LVA

  • E. coli

LVA

  • E. coli

LVA

  • E. coli

LVA

  • E. coli

Sunday, October 9, 2011

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SLIDE 22

2011

Design Fabrication Analysis

Characterization Characterization

Parts used

Sunday, October 9, 2011

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SLIDE 23

2011

Design Fabrication Analysis

Characterization Characterization

Parts used

Fluorescent Proteins

Sunday, October 9, 2011

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SLIDE 24

2011

Design Fabrication Analysis

Characterization Characterization

Parts used

mCitrine

AcGFP

Fluorescent Proteins

tagRFP

ECFP

mCherry

mTFP1

GFP

mOrange

mRuby

tdTomato

EYFP

YPet

Venus

CFP tGFP sfGFP DsRed

Sunday, October 9, 2011

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SLIDE 25

2011

Design Fabrication Analysis

Characterization Characterization

Parts used

mCitrine

AcGFP

Fluorescent Proteins Degradation Tags

tagRFP

ECFP

mCherry

mTFP1

Sul20C LVA PEST Ubi-Lys

GFP

mOrange

mRuby

tdTomato

EYFP

YPet

Venus

CFP tGFP sfGFP DsRed

No Tag No Tag

Sunday, October 9, 2011

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SLIDE 26

2011

Design Fabrication Analysis

Characterization Characterization

Parts used

pLac

mCitrine

AcGFP

pGal Fluorescent Proteins Degradation Tags Plasmids/ Promoters

tagRFP

ECFP

mCherry

mTFP1

Sul20C LVA PEST Ubi-Lys

GFP

mOrange

mRuby

tdTomato

EYFP

YPet

Venus

CFP tGFP sfGFP DsRed

No Tag No Tag

Sunday, October 9, 2011

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SLIDE 27

2011

Design Fabrication Analysis

Characterization Characterization

Parts used

pLac

mCitrine

AcGFP

  • E. coli
  • S. cerevisiae

pGal Fluorescent Proteins Degradation Tags Plasmids/ Promoters Organisms

tagRFP

ECFP

mCherry

mTFP1

Sul20C LVA PEST Ubi-Lys

GFP

mOrange

mRuby

tdTomato

EYFP

YPet

Venus

http://www.mansfieldct.org/schools/mms/staff/hand/Bactvsvir.htm Credit: Visuals Unlimited Inc Photographer: Thomas Deernick

CFP tGFP sfGFP DsRed

No Tag No Tag

Sunday, October 9, 2011

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SLIDE 28

2011

Design Fabrication Analysis

Characterization Characterization

GenoCAD

Sunday, October 9, 2011

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SLIDE 29

2011

Design Fabrication Analysis

Characterization Characterization

GenoCAD

  • Parts collected - how to put them

together, not just physically but digitally

Sunday, October 9, 2011

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SLIDE 30

2011

Design Fabrication Analysis

Characterization Characterization

GenoCAD

  • Parts collected - how to put them

together, not just physically but digitally

Sunday, October 9, 2011

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SLIDE 31

2011

Design Fabrication Analysis

Characterization Characterization

GenoCAD

  • Parts collected - how to put them

together, not just physically but digitally

Sunday, October 9, 2011

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SLIDE 32

2011

Design Fabrication Analysis

Characterization Characterization

GenoCAD

  • Parts collected - how to put them

together, not just physically but digitally

  • Grammar helps us put parts together,

and attributes associate meaning with parts, which we’ll use for modeling

Sunday, October 9, 2011

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SLIDE 33

2011

Design Fabrication Analysis

Characterization Characterization

GenoCAD

  • Parts collected - how to put them

together, not just physically but digitally

  • Grammar helps us put parts together,

and attributes associate meaning with parts, which we’ll use for modeling

  • Created custom grammar for this

project - we’ll focus on the grammar now and attributes later

Sunday, October 9, 2011

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SLIDE 34

2011

Design Fabrication Analysis

Characterization Characterization

Our Grammar - It’s Big

  • E. coli Insert

Suf FP Single FP (Tag) Fusion FP (Link) FP Pre Pro Vector Stop Reporter Single FP Fusion

  • E. coli Construct

Suffix

  • S. cerevisiae Insert

Stop Reporter Single FP Fusion Promoter Prefix

  • S. cerevisiae Construct

Single FP Fusion

Start

Stop Stop

Stop

Stop p

(Tag) (Tag) (Tag) Suf FP (Tag) FP (Link) FP Pre Pro Stop (Tag) (Tag) (Tag) Suffix Stop Reporter Promoter Prefix Vec FP (Tag) FP (Link) FP Stop (Tag) (Tag) (Tag) Vec FP (Tag) FP (Link) FP Stop (Tag) (Tag) (Tag) Vector Stop Reporter

Sunday, October 9, 2011

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SLIDE 35

2011

Design Fabrication Analysis

Characterization Characterization

Our Grammar - It’s Big

  • E. coli Insert

Suf FP Single FP (Tag) Fusion FP (Link) FP Pre Pro Vector Stop Reporter Single FP Fusion

  • E. coli Construct

Suffix

  • S. cerevisiae Insert

Stop Reporter Single FP Fusion Promoter Prefix

  • S. cerevisiae Construct

Single FP Fusion

Start

Stop Stop

Stop

Stop p

(Tag) (Tag) (Tag) Suf FP (Tag) FP (Link) FP Pre Pro Stop (Tag) (Tag) (Tag) Suffix Stop Reporter Promoter Prefix Vec FP (Tag) FP (Link) FP Stop (Tag) (Tag) (Tag) Vec FP (Tag) FP (Link) FP Stop (Tag) (Tag) (Tag) Vector Stop Reporter

Sunday, October 9, 2011

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SLIDE 36

2011

Design Fabrication Analysis

Characterization Characterization

Our Grammar - Example

Vector

  • E. coli Construct

Vector Reporter FP Single FP (Tag) (Tag) Fusion FP (Tag) (Tag) (Link) FP

Goal: Design a fluorescent reporter with one fluorescent protein, one degradation tag, in E. coli-compatible vector

Sunday, October 9, 2011

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SLIDE 37

2011

Design Fabrication Analysis

Characterization Characterization

Our Grammar - Example

Vector

  • E. coli Construct

Specific Vector Reporter FP Single FP (Tag) (Tag) Fusion FP (Tag) (Tag) (Link) FP

Goal: Design a fluorescent reporter with one fluorescent protein, one degradation tag, in E. coli-compatible vector

Sunday, October 9, 2011

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SLIDE 38

2011

Design Fabrication Analysis

Characterization Characterization

Our Grammar - Example

Vector

  • E. coli Construct

Reporter FP Single FP (Tag) (Tag) Fusion FP (Tag) (Tag) (Link) FP

Goal: Design a fluorescent reporter with one fluorescent protein, one degradation tag, in E. coli-compatible vector

Specific Vector

Sunday, October 9, 2011

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SLIDE 39

2011

Design Fabrication Analysis

Characterization Characterization

Our Grammar - Example

Vector

  • E. coli Construct

Reporter

Specific FP

Single FP (Tag)

Specific Tag

Fusion FP (Tag) (Tag) (Link) FP

Goal: Design a fluorescent reporter with one fluorescent protein, one degradation tag, in E. coli-compatible vector

Specific Vector

Sunday, October 9, 2011

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SLIDE 40

2011

Design Fabrication Analysis

Characterization Characterization

Roadmap - Design

Name Tag Organism Design Cloning Screening Sequencing Imaging Data Analysis

AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine None

  • S. cerevisiae

X None

  • S. cerevisiae

X None

  • S. cerevisiae

X None

  • S. cerevisiae

X None

  • S. cerevisiae

X None

  • S. cerevisiae

X None

  • E. coli

X None

  • E. coli

X None

  • E. coli

X None

  • E. coli

X None

  • E. coli

X PEST

  • S. cerevisiae

X PEST

  • S. cerevisiae

X PEST

  • S. cerevisiae

X PEST

  • S. cerevisiae

X PEST

  • S. cerevisiae

X PEST

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X

Sunday, October 9, 2011

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SLIDE 41

2011

Design Fabrication Analysis

Characterization

Roadmap - Fabrication

Name Tag Organism Design Cloning Screening Sequencing Imaging Data Analysis

AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X None

  • E. coli

X X X X None

  • E. coli

X X X X None

  • E. coli

X X X X None

  • E. coli

X X X X None

  • E. coli

X X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X LVA

  • E. coli

X X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X

Sunday, October 9, 2011

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SLIDE 42

2011

Design Fabrication Analysis

Characterization

Fluorescence Microscopy

  • Automated microfluidics
  • GenoSIGHT - custom

control software

  • Tracks hundreds of

individual cells

  • Optimized time resolution

Sunday, October 9, 2011

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SLIDE 43

2011

Design Fabrication Analysis

Characterization

Microscopy - Stabilization

Sunday, October 9, 2011

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SLIDE 44

2011

Design Fabrication Analysis

Characterization

Microscopy - Stabilization

Sunday, October 9, 2011

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SLIDE 45

2011

Design Fabrication Analysis

Characterization

Microscopy - Stabilization

Sunday, October 9, 2011

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SLIDE 46

2011

Design Fabrication Analysis

Characterization

Microscopy - Induction

Sunday, October 9, 2011

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SLIDE 47

2011

Design Fabrication Analysis

Characterization

Microscopy - Induction

Sunday, October 9, 2011

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SLIDE 48

2011

Design Fabrication Analysis

Characterization

Microscopy - Inhibition

Sunday, October 9, 2011

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SLIDE 49

2011

Design Fabrication Analysis

Characterization

Microscopy - Inhibition

Sunday, October 9, 2011

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SLIDE 50

2011

Design Fabrication Analysis

Characterization

Roadmap - Imaging

Name Tag Organism Design Cloning Screening Sequencing Imaging Data Analysis

AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • E. coli

X X X X X None

  • E. coli

X X X X X None

  • E. coli

X X X X X None

  • E. coli

X X X X X None

  • E. coli

X X X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X LVA

  • E. coli

X X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X

Sunday, October 9, 2011

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SLIDE 51

2011

Design Fabrication Analysis

Characterization

Modeling

degraded

Immature Protein Mature protein

expression degraded

Sunday, October 9, 2011

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SLIDE 52

2011

Design Fabrication Analysis

Characterization

Modeling

d

degraded

Immature Protein Mature protein

m

expression

d

degraded

Sunday, October 9, 2011

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SLIDE 53

2011

Design Fabrication Analysis

Characterization

Modeling

d

degraded

Immature Protein Mature protein

m

expression

d

degraded

Sunday, October 9, 2011

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SLIDE 54

2011

Design Fabrication Analysis

Characterization

Modeling

d

degraded

Immature Protein Mature protein

m

expression

d

degraded

FP Fluorescent Reporter (Tag) (Tag)

m, d d d

Sunday, October 9, 2011

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SLIDE 55

2011

Design Fabrication Analysis

Characterization

Modeling

d

degraded

Immature Protein Mature protein

m

expression

d

degraded

FP Fluorescent Reporter (Tag) (Tag)

m, d d d

Sunday, October 9, 2011

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SLIDE 56

2011

Design Fabrication Analysis

Characterization

Modeling

d

degraded

Immature Protein Mature protein

m

expression

d

degraded

FP Fluorescent Reporter (Tag) (Tag)

Time F l u

  • r

e s c e n c e

m, d d d

Sunday, October 9, 2011

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SLIDE 57

2011

Design Fabrication Analysis

Characterization

Parameter Estimation

fluorescence over time for one cell

Sunday, October 9, 2011

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SLIDE 58

2011

Design Fabrication Analysis

Characterization

Parameter Estimation

R-squared = 0.2

Sunday, October 9, 2011

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SLIDE 59

2011

Design Fabrication Analysis

Characterization

Parameter Estimation

R-squared = 0.5

Sunday, October 9, 2011

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SLIDE 60

2011

Design Fabrication Analysis

Characterization

Parameter Estimation

Sunday, October 9, 2011

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SLIDE 61

2011

Design Fabrication Analysis

Characterization

Parameter Estimation

Sunday, October 9, 2011

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SLIDE 62

2011

Design Fabrication Analysis

Characterization

Methods of Data Processing

F l u

  • r

e s c e n c e Time

Sunday, October 9, 2011

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SLIDE 63

2011

Design Fabrication Analysis

Characterization

Methods of Data Processing

F l u

  • r

e s c e n c e Time

Sunday, October 9, 2011

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SLIDE 64

2011

Design Fabrication Analysis

Characterization

Methods of Data Processing

“Cell-by-Cell” Method

F l u

  • r

e s c e n c e Time

Time Fluorescence

Sunday, October 9, 2011

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SLIDE 65

2011

Design Fabrication Analysis

Characterization

Methods of Data Processing

“Averaging” Method “Cell-by-Cell” Method

F l u

  • r

e s c e n c e Time

Fluorescence Time Time Fluorescence

Sunday, October 9, 2011

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SLIDE 66

2011

Design Fabrication Analysis

Characterization

Distribution of Data - mCherry

m = 0.0161 mol/min d = 0.0045 mol/min R^2 = .4

Sunday, October 9, 2011

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SLIDE 67

2011

Design Fabrication Analysis

Characterization

Example Curve

Time F l u

  • r

e s c e n c e

Sunday, October 9, 2011

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SLIDE 68

2011

Design Fabrication Analysis

Characterization

Roadmap - Analysis

Name Tag Organism Design Cloning Screening Sequencing Imaging Data Analysis

AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCherry mCitrine AcGFP ECFP GFPmut3b mCherry(E2060) mCitrine None

  • S. cerevisiae

X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • S. cerevisiae

X X X X X None

  • E. coli

X X X X X None

  • E. coli

X X X X X None

  • E. coli

X X X X X X None

  • E. coli

X X X X X X None

  • E. coli

X X X X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X PEST

  • S. cerevisiae

X X X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X Ubi-Lys

  • S. cerevisiae

X LVA

  • E. coli

X X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X LVA

  • E. coli

X

Sunday, October 9, 2011

slide-69
SLIDE 69

2011

Design Fabrication Analysis

Characterization

Comparing results

Gene Organism m

Maturation Half Time

d

Degradation Half Time

mCitrine

  • E. coli

0.0029 ±10-5 239 min 0.0031 ±10-5 233 min GFPmut3b

  • E. coli

0.034 ±10-4 20.4 min** 0.0076 ±10-5 91.2 min*** mCherry

  • E. coli

0.0161 ±10-5 43 min* 0.0046 ±10-5 151 min

fast degradation could be due to photobleaching *agrees with in vitro literature value (15 min, Shaner 2004) ***disagrees with 2008 KDLeuven iGEM results (1980 minutes) **agrees with in vivo literature value for GFPmut3 (6 min, Megerle 2008)

Sunday, October 9, 2011

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SLIDE 70

2011

Design Fabrication Analysis

Characterization

Comparing Results

Sunday, October 9, 2011

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SLIDE 71

2011

Design Fabrication Analysis

Characterization

Challenges of Characterization

F l u

  • r

e s c e n c e Time F l u

  • r

e s c e n c e Time

  • Increasing light exposure increases degradation rate
  • Linear relationship - 2x light leads to 2x degradation

Sunday, October 9, 2011

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SLIDE 72

2011 Conclusion

Sunday, October 9, 2011

slide-73
SLIDE 73

2011 Conclusion

  • 1. Characterization is challenging

Sunday, October 9, 2011

slide-74
SLIDE 74

2011 Conclusion

  • 1. Characterization is challenging
  • 2. We can obtain values consistent with literature

Sunday, October 9, 2011

slide-75
SLIDE 75

2011 Conclusion

  • 1. Characterization is challenging
  • 2. We can obtain values consistent with literature
  • 3. Our workflow and tools can increase throughput

Sunday, October 9, 2011

slide-76
SLIDE 76

2011 Achievements

Parts submitted Work Completed

Sunday, October 9, 2011

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SLIDE 77

2011 Acknowledgements

The Team would like to thank Dr. Jean Peccoud, Laura Adam, Dr. David Ball, Dr. Eborall, Matthew Lux and Julie Marchand for their guidance in the execution of the project. Funding and support:

Sunday, October 9, 2011

slide-78
SLIDE 78

2011 Citations

Elowitz MB, Leibler SA. Synthetic Oscillatory Network of Transcriptional Regulators. Nature. 2000 Jan 20;403(6767):335-8. Shaner NC, Campbell RE, Steinbach PA, Giepmans BG, Palmer AE, Tsien RY. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature Biotechnology. 2004 Dec; 22(12):1567-1572 Megerle JA, Fritz G, Gerland U, Jung K, Rädler JO. Timing and dynamics of single cell gene expression in the arabinose utilization

  • system. Biophys J. 2008 Aug95(4):2103-15.

Gordon, A. Colman-Lerner, T.E. Chin, K.R. Benjamin, R.C. Yu and

  • R. Brent, Single-cell quantification of molecules and rates using
  • pen-source microscope-based cytometry. Nat.

Methods, 4 (2007), pp. 175–181.

Sunday, October 9, 2011

slide-79
SLIDE 79

2011 End

Sunday, October 9, 2011