Difference between revisions of "Part:BBa K117002:Experience"

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<I>Tokyo Tech 2011</I>
 
<I>Tokyo Tech 2011</I>
 
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[[Image:PlsrA_activity.png|thumb|center|700px|Median fluorescence intensity(MFI) of BBa_K649104 was much higher than that of promoterless-gfp(negative control).<br>This work is done by Takuya Tsubaki.]]
  
 
This part(BBa_K117002) does not work properly. To confirm this, we introduced a gfp gene(BBa_J54103) downstream of the promoter. As a consequence, fluorescence intensity of promoter lsrA(BBa_K117002)-gfp was almost the same as promoterless-gfp(negative control), showing that promoter lsrA(BBa_K117002) does not work properly.  
 
This part(BBa_K117002) does not work properly. To confirm this, we introduced a gfp gene(BBa_J54103) downstream of the promoter. As a consequence, fluorescence intensity of promoter lsrA(BBa_K117002)-gfp was almost the same as promoterless-gfp(negative control), showing that promoter lsrA(BBa_K117002) does not work properly.  

Revision as of 10:58, 3 October 2011

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K117002

This promoter is activated indirectly by AI-2 to promote whatever downstream gene ligated behind it.

Characterisation:

For information on characterisation of this new part, please visit Part K117010 Experience and Part K117008 Experience

User Reviews

UNIQbbecabff348a414e-partinfo-00000000-QINU UNIQbbecabff348a414e-partinfo-00000001-QINU

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Tokyo Tech 2011

Median fluorescence intensity(MFI) of BBa_K649104 was much higher than that of promoterless-gfp(negative control).
This work is done by Takuya Tsubaki.

This part(BBa_K117002) does not work properly. To confirm this, we introduced a gfp gene(BBa_J54103) downstream of the promoter. As a consequence, fluorescence intensity of promoter lsrA(BBa_K117002)-gfp was almost the same as promoterless-gfp(negative control), showing that promoter lsrA(BBa_K117002) does not work properly.


We improved this part.Our new promoter lsrA(BBa_K649100) works.


If you want to know why our part works, click [here] and visit our team's wiki.

[sample]

pSB1A2 Ptet-gfp(JD22597)

pSB6A1 promoterless-gfp(JD22597)

pSB1A2 PlsrA-gfp(BBa_K649104)(JD22597)

pSB1A2 PlsrA-gfp(BBa_K11702-GFP)(JD22597)

[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 1:100.

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.



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