Difference between revisions of "Part:BBa K2548001"

Revision as of 11:23, 30 September 2018 (view source) Registry (Talk | contribs)		Latest revision as of 21:05, 17 October 2018 (view source) Joycelalala (Talk | contribs)
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	<partinfo>BBa_K2548001 short</partinfo>		<partinfo>BBa_K2548001 short</partinfo>
		+	<p>RhlR coding device with 60 times of predicted RBS strength compared with BBa_K2548000. The RBS sequence is obtained from Salis Lab RBS Calculator V2.0.</p>
−	RhlR coding device with 60 times of predicted RBS strength compared with BBa_K2548000. The RBS sequence is obtained from Salis Lab RBS Calculator V2.0.	+	<h2>Construction of A. hydrophila Sensor Device III (BBa_K2548001 + BBa_K2548003) </h2>
		+	https://static.igem.org/mediawiki/parts/thumb/b/b7/Sensordevice3.png/800px-Sensordevice3.png
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		+	<h2>Characterization of <i>A. hydrophila</i>Sensor Device III </h2>
		+	<p>We calculated four new RBS sequence with strength of 10, 20, 30, and 60 times of BBa_B0034 and assembled them with our sensor device. We tested the GFP production rate of these four devices, and found the device with 60 times of RBS strength gave the best result. The GFP production rate of this device at 9.90E-8 M of C4-HSL is 2.015 per cell RFU OD-1 min-1:</p>
		+	[[Image:GFP-ORI.png|thumb|center|400px|Fig.1 GFP production rate of sensor device III]]<br>
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		+	<p>This time, the difference between GFP production rate at 0 M and 9.90E-8 M of C4-HSL reached 35.7% of the expression level without induction, enabling people to detect <i>A. hydrophila</i> more easily from fish feces by using fluorescence detector. Moreover, as Mr. Zhang predicted before, the cooperativity between RhlR and C4-HSL was improved. Figure x. shows the modelling results of the three sensor devices. The slope of device III is the steepest one, and the Hill coefficient of the function, which describes the cooperativity between inducer and RhlR, increased from 1.154 in device I to 2.250 in this device.</p>
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		+	[[Image:60ORImodel.png|thumb|center|400px|Fig.2 Modelling results of three A. hydrophila sensor devices]]<br>
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Latest revision as of 21:05, 17 October 2018

Constitutive promoter- RBS - rhlR (LVA tagged) -terminator

RhlR coding device with 60 times of predicted RBS strength compared with BBa_K2548000. The RBS sequence is obtained from Salis Lab RBS Calculator V2.0.

Construction of A. hydrophila Sensor Device III (BBa_K2548001 + BBa_K2548003)

Characterization of A. hydrophilaSensor Device III

We calculated four new RBS sequence with strength of 10, 20, 30, and 60 times of BBa_B0034 and assembled them with our sensor device. We tested the GFP production rate of these four devices, and found the device with 60 times of RBS strength gave the best result. The GFP production rate of this device at 9.90E-8 M of C4-HSL is 2.015 per cell RFU OD-1 min-1:

This time, the difference between GFP production rate at 0 M and 9.90E-8 M of C4-HSL reached 35.7% of the expression level without induction, enabling people to detect A. hydrophila more easily from fish feces by using fluorescence detector. Moreover, as Mr. Zhang predicted before, the cooperativity between RhlR and C4-HSL was improved. Figure x. shows the modelling results of the three sensor devices. The slope of device III is the steepest one, and the Hill coefficient of the function, which describes the cooperativity between inducer and RhlR, increased from 1.154 in device I to 2.250 in this device.

Sequence and Features