Top Banner
Mechanism & Function A transcription factor (LuxR) that is active in the presence of a cell-cell signaling molecule (3OC6HSL) is controlled by a regulated operator (PLtetO-1). Device input is 3OC6HSL. Device output is PoPS from a LuxR-regulated operator. If used in a cell containing TetR then a second input such as aTc can be used to produce a Boolean AND function. BBa_F2620 3OC 6 HSL PoPS Receiver Registry of Standard Biological Parts making life better, one part at a time License: Public Conditions (abridged) Output: PoPS measured via BBa_E0240 Culture: Supplemented M9, 37ºC Plasmid: pSB3K3 Chassis: MG1655 *Equipment: PE Victor3 multi-well fluorimeter **Equipment: BD FACScan cytometer 1E0 1E1 1E2 1E3 1E4 GFP (arbitrary units) Doublings 20 38 56 74 92 High Input (1E -7 M 3OC 6 HSL) 92 1E0 1E1 1E2 1E3 1E4 Doublings 20 38 56 74 Low Input (0 M 3OC 6 HSL) GFP (arbitrary units) Reliability** Genetic: >92/>56 culture doublings Performance: >92/>56 culture doublings (low/high input during propagation) Transcriptional Output Demand (low/high input) Nucleotides: 0 / 6xNt nucleotides cell -1 s -1 Polymerases: 0 / 1.5E-1xNt RNAP cell -1 (Nt = downstream transcript length) Chassis: MC4100, MG1655, and DH5 Plasmids: pSB3K3 and pSB1A2 Devices: E0240, E0430 and E0434 BBa_F2620 Response Time: <1 min BBa_T9002 Response Time: 6±1 min Inputs: 0 M (Low), 1E-07 M (High) 3OC 6 HSL P max : 6.6 PoPS cell -1 K: 1.5E-09 M 3OC 6 HSL n: 1.6 Static Performance* P out = P max [3OC 6 HSL] n K n + [3OC 6 HSL] n 0E+00 1E−10 1E−09 1E−08 1E−07 1E−06 1E−05 1E−04 0 100 200 300 400 500 600 [3OC 6 HSL] (M) GFP synthesis rate (molecules cell −1 s −1 ) Population Mean Colony Range Hill Equation 0 1 2 3 4 5 6 7 8 PoPS cell −1 Part Compatibility (qualitative) Authors: Barry Canton Ania Labno Updated: March 2008 Signaling Devices http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620 R0040 B0034 C0062 B0015 R0062 PLtetO-1 RBS luxR Term. Plux,R Component Parts −10 0 10 20 30 40 50 0 100 200 300 400 500 600 + 3OC 6 HSL Time (min) GFP synthesis rate (molecules cell −1 s −1 ) GFP synthesis rate (Low Input) GFP synthesis rate (High Input) Polynomial Fit (High Input) PoPS (High Input) 0 1 2 3 4 5 6 7 8 PoPS cell −1 0E+00 1E−10 1E−09 1E−08 1E−07 1E−06 1E−05 1E−04 0 100 200 300 400 500 600 [AHL] (M) GFP synthesis rate (molecules cell −1 s −1 ) C 4 HSL C 6 HSL 3OC 6 HSL C 7 HSL C 8 HSL 3OC 8 HSL C 10 HSL C 12 HSL 0 1 2 3 4 5 6 7 8 PoPS cell −1 Input Compatibility* Dynamic Performance*
1

BBa F2620 HSL PoPS Receiver - parts.igem.orgparts.igem.org/wiki/images/f/fe/EndyFig1-F2620DataSheet.pdf · (Nt = downstream transcript length) Chassis: MC4100, MG1655, and DH5 Plasmids:

Aug 05, 2020

Download

Documents

dariahiddleston
Welcome message from author
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Page 1: BBa F2620 HSL PoPS Receiver - parts.igem.orgparts.igem.org/wiki/images/f/fe/EndyFig1-F2620DataSheet.pdf · (Nt = downstream transcript length) Chassis: MC4100, MG1655, and DH5 Plasmids:

Mechanism & FunctionA transcription factor (LuxR) that is active in the presence of a cell-cell signaling molecule (3OC6HSL) is controlled by a regulated operator (PLtetO-1). Device input is 3OC6HSL. Device output is PoPS from a LuxR-regulated operator. If used in a cell containing TetR then a second input such as aTc can be used to produce a Boolean AND function.

BBa_F26203OC6HSL PoPS Receiver

Registry of Standard Biological Parts making life better, one part at a time

License: Public

Conditions (abridged)Output: PoPS measured via BBa_E0240Culture: Supplemented M9, 37ºCPlasmid: pSB3K3Chassis: MG1655*Equipment: PE Victor3 multi-well fluorimeter**Equipment: BD FACScan cytometer

1E01E1

1E21E3

1E4

GFP (arbitrary units)

Dou

blin

gs

2038

56

74

92

High Input (1E -7 M 3OC6HSL)

92

1E01E1

1E21E3

1E4

Dou

blin

gs

2038

56

74

Low Input(0 M 3OC6HSL)

GFP (arbitrary units)

Reliability**

Genetic: >92/>56 culture doublingsPerformance: >92/>56 culture doublings (low/high input during propagation)

Transcriptional Output Demand (low/high input)Nucleotides: 0 / 6xNt nucleotides cell-1 s-1

Polymerases: 0 / 1.5E-1xNt RNAP cell-1

(Nt = downstream transcript length)

Chassis: MC4100, MG1655, and DH5Plasmids: pSB3K3 and pSB1A2Devices: E0240, E0430 and E0434

BBa_F2620 Response Time: <1 minBBa_T9002 Response Time: 6±1 minInputs: 0 M (Low), 1E-07 M (High) 3OC6HSL

Pmax: 6.6 PoPS cell-1 K: 1.5E-09 M 3OC6HSLn: 1.6

Static Performance*

Pout =Pmax [3OC6HSL]n

Kn + [3OC6HSL]n

0E+00 1E−10 1E−09 1E−08 1E−07 1E−06 1E−05 1E−040

100

200

300

400

500

600

[3OC6HSL] (M)

GFP

syn

thes

is ra

te (m

olec

ules

cel

l−1 s−1

)

Population MeanColony RangeHill Equation

0

1

2

3

4

5

6

7

8

PoPS

cel

l−1

Part Compatibility (qualitative)

Authors: Barry Canton Ania Labno

Updated: March 2008

Sig

nal

ing

Dev

ices

http

://pa

rts.

mit.

edu/

regi

stry

/inde

x.ph

p/P

art:B

Ba_

F26

20

BBa_F2620

R0040 B0034 C0062 B0015 R0062 PLtetO-1 RBS luxR Term. Plux,R

Component Parts

−10 0 10 20 30 40 500

100

200

300

400

500

600

+ 3OC6HSL

Time (min)

GFP

syn

thes

is ra

te (m

olec

ules

cel

l−1 s−1

)

GFP synthesis rate (Low Input)GFP synthesis rate (High Input)Polynomial Fit (High Input)PoPS (High Input)

0

1

2

3

4

5

6

7

8

PoPS

cel

l−1

0E+00 1E−10 1E−09 1E−08 1E−07 1E−06 1E−05 1E−040

100

200

300

400

500

600

[AHL] (M)

GFP

syn

thes

is ra

te (m

olec

ules

cel

l−1 s−1

)

C4HSLC6HSL3OC6HSLC7HSLC8HSL3OC8HSLC10HSLC12HSL

0

1

2

3

4

5

6

7

8

PoPS

cel

l−1

Input Compatibility*

Dynamic Performance*