Difference between revisions of "Part:BBa K082026:Experience"

(→‎User Reviews)
(→‎User Reviews)
Line 29: Line 29:
 
<I>Tokyo Tech iGEM2010</I>
 
<I>Tokyo Tech iGEM2010</I>
 
|width='60%' valign='top'|
 
|width='60%' valign='top'|
 +
Since sequence data of BBa_K082026 is incorrect, please refer to [https://parts.igem.org/wiki/index.php/Part:BBa_K395400 BBa_K395400].
 +
However, we confirmed that both BBa_K395400 and BBa_K082026 have the same activity.
 +
<br>
 +
In order to characterize K395400, LacI Mutant, we constructed BBa_K395401 combining I0500 and K082026(K395400) on pSB1A3 and used BBa_I20260 as a positive control and used promoterless gfp on pSB3K3 as a negative control.
 +
Furthermore, we constructed BBa_K395402 combining I0500 and BBa_I 732820 on pSB1A3 as a control experiment.
 +
As a GFP reporter, we used BBa_I7106 on pSB3K3.
 +
In order to measure the function of K395400), we introduced BBa_I7106 (lacI+pL-rbs-GFP-ter) on pSB3K3 and BBa_K395401/BBa_K395402 into DH5α. <br>
 +
<br>We confirmed that product of lacIM1 shows weaker repression to lac promoter than its wild type.<br>
 +
For more information, see [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 our work in Tokyo_Tech 2010 wiki]
  
−
[[Image:Tokyotech_LacIM1_system.png|left|thumb|200px|Figure3-2-1. ]]
 
−
[[Image:Tokyotech_LacIM1_data.png|center|thumb|300px|Figure3-2-2. Repression efficiency of LacIM1 (BBa_K395401) / LacIWT (BBa_K395402) exposed to arabinose and IPTG.  This work is done by Mitsuhiko Odera ]]
 
−
 
−
LacIM1 shows weaker repression than LacIWT because it has a lower affinity to lac promoter [1].  Though this part was registered by USTC(2008), it was not well characterized in their wiki.  Therefore, we characterized this part.
 
−
 
−
 
−
We confirmed that lacIM1 shows weaker repression than LacIWT.
 
−
 
−
(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])
 
  
  

Revision as of 20:56, 27 October 2010

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_K082026

User Reviews

UNIQa11caab84c754993-partinfo-00000000-QINU UNIQa11caab84c754993-partinfo-00000001-QINU


UNIQa11caab84c754993-partinfo-00000002-QINU

Characterization of BBa_K082026

Tokyo Tech iGEM2010

Since sequence data of BBa_K082026 is incorrect, please refer to BBa_K395400. However, we confirmed that both BBa_K395400 and BBa_K082026 have the same activity.
In order to characterize K395400, LacI Mutant, we constructed BBa_K395401 combining I0500 and K082026(K395400) on pSB1A3 and used BBa_I20260 as a positive control and used promoterless gfp on pSB3K3 as a negative control. Furthermore, we constructed BBa_K395402 combining I0500 and BBa_I 732820 on pSB1A3 as a control experiment. As a GFP reporter, we used BBa_I7106 on pSB3K3. In order to measure the function of K395400), we introduced BBa_I7106 (lacI+pL-rbs-GFP-ter) on pSB3K3 and BBa_K395401/BBa_K395402 into DH5α.

We confirmed that product of lacIM1 shows weaker repression to lac promoter than its wild type.
For more information, see [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 our work in Tokyo_Tech 2010 wiki]


;

UNIQa11caab84c754993-partinfo-00000003-QINU