Difference between revisions of "Part:BBa K649105:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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===Applications of BBa_K649105===
 
===Applications of BBa_K649105===
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[[Image:LsrR repression.png|thumb|center|300px|Median fluorescence intensity(MFI) is decreased by LsrR repression.<br>This work is done by Hiroki Yoshise.]]
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We confirmed that LsrR represses lsrA promoter.For this purpose, we constructed BBa_K649105(PlsrA-gfp-PlsrR-lsrR) and compared fluorescence intensity levels of BBa_K649104(PlsrA-RBS-gfp) and BBa_K649105.As a consequence, Median fluorescence intensity(MFI) of BBa_K649104(PlsrA-RBS-gfp) is 3-fold higher than that of BBa_K649105(PlsrA-gfp-PlsrR-lsrR).  This result shows that LsrR represses lsrA promoter and  BBa_K649105 works.
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[Sample]
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pSB6A1 Ptet-GFP RBS1-12(JM2.300)(positive control)
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pSB6A1 ⊿P-GFP(JM2.300)(negative control)
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pSB3K3 PlsrA-GFP(MG1655)
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pSB3K3 PlsrA-GFP-PlsrR-lsrR(MG1655)
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[Method]
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1. Overnight cultures of reporter strains grown at 37 °C in LB medium containing appropriate antibiotics were diluted 1:100 into 3 ml of LB medium and were incubated at 37 °C as fresh cultures.
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2. After their OD590 reached 0.15, the fresh cultures were diluted 1:10 or 1:100.
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3. After 4-hour incubation at 37 °C, 1 ml of each culture was moved to 1.6ml tube and its fluorescence intensity was measured with a flow cytometer.
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===User Reviews===
 
===User Reviews===

Revision as of 07:23, 2 October 2011

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Please enter how you used this part and how it worked out.

Applications of BBa_K649105

Median fluorescence intensity(MFI) is decreased by LsrR repression.
This work is done by Hiroki Yoshise.


We confirmed that LsrR represses lsrA promoter.For this purpose, we constructed BBa_K649105(PlsrA-gfp-PlsrR-lsrR) and compared fluorescence intensity levels of BBa_K649104(PlsrA-RBS-gfp) and BBa_K649105.As a consequence, Median fluorescence intensity(MFI) of BBa_K649104(PlsrA-RBS-gfp) is 3-fold higher than that of BBa_K649105(PlsrA-gfp-PlsrR-lsrR). This result shows that LsrR represses lsrA promoter and BBa_K649105 works.

[Sample] pSB6A1 Ptet-GFP RBS1-12(JM2.300)(positive control) pSB6A1 ⊿P-GFP(JM2.300)(negative control) pSB3K3 PlsrA-GFP(MG1655) pSB3K3 PlsrA-GFP-PlsrR-lsrR(MG1655)

[Method] 1. Overnight cultures of reporter strains grown at 37 °C in LB medium containing appropriate antibiotics were diluted 1:100 into 3 ml of LB medium and were incubated at 37 °C as fresh cultures. 2. After their OD590 reached 0.15, the fresh cultures were diluted 1:10 or 1:100. 3. After 4-hour incubation at 37 °C, 1 ml of each culture was moved to 1.6ml tube and its fluorescence intensity was measured with a flow cytometer.



User Reviews

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