Difference between revisions of "Part:BBa K2407013"

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	<partinfo>BBa_K2407013 short</partinfo>		<partinfo>BBa_K2407013 short</partinfo>
−	loading...	+	<h3>Design</h3>
		+	<p>This part was improved from <partinfo>BBa_K2407000</partinfo> and the source can be tracked back to <partinfo>BBa_K2165004</partinfo>.</p>
		+
		+	<p>In our experiment, we noticed that <i>CUP1</i> promoter still has a certain degree of leakage expression. To make a better biosensor, we planned to reduce the leakage expression and increase the sensitivity. To reach this goal, we took the fluorescence intensity at both induction or not into evaluation indexes.</p>
		+
		+	<p>The technology of error-prone PCR was taken into our experiment. Although there are many methods to introduce genetic diversity into a parent sequence, error-prone PCR is the most common way of creating a combinatorial library based on a single sequence. By adding some heavy metal ions into the PCR buffer and preparing dNTPs with different composition, new mutants were introduced into <i>CUP1</i> promoter.</p>
		+
		+	<p>This is one of the mutants from <i>CUP1</i> promoter, which was named with EP-3. The changes in sequence were uploaded in the "sequence and feature" part.
		+
		+	<h3>Characterization</h3>
		+	<p>We tested the leakage expression and response rages of each selected mutants, and results were shown below.</p>
		+
		+	[[File:Tianjin-Demonstrate-4-5-leakage.png|700px|thumb|center|'''Fig 1'''.The leakage expression of different promoters ]]
		+
		+	[[File:Tianjin-Demonstrate-4-6-EP.png|700px|thumb|center|'''Fig 1'''.Different biosensors showed different sensitivities and response rages ]]
		+
	<!-- Add more about the biology of this part here		<!-- Add more about the biology of this part here

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Revision as of 12:06, 27 October 2017

a modified CUP1 promoter obtained by Error-Prone PCR EP3

Design

This part was improved from BBa_K2407000 and the source can be tracked back to BBa_K2165004.

In our experiment, we noticed that CUP1 promoter still has a certain degree of leakage expression. To make a better biosensor, we planned to reduce the leakage expression and increase the sensitivity. To reach this goal, we took the fluorescence intensity at both induction or not into evaluation indexes.

The technology of error-prone PCR was taken into our experiment. Although there are many methods to introduce genetic diversity into a parent sequence, error-prone PCR is the most common way of creating a combinatorial library based on a single sequence. By adding some heavy metal ions into the PCR buffer and preparing dNTPs with different composition, new mutants were introduced into CUP1 promoter.

This is one of the mutants from CUP1 promoter, which was named with EP-3. The changes in sequence were uploaded in the "sequence and feature" part.

Characterization

<p>We tested the leakage expression and response rages of each selected mutants, and results were shown below.

Sequence and Features