Difference between revisions of "Part:BBa K395400"

Revision as of 20:33, 27 October 2010

RBS-lacIM1-ter

Though LacIM1(BBa_K082026) has been registered, sequence data of BBa_K082026 is incorrect. Therefore, we resistered this part again.
LacIM1 shows weaker repression than LacIWT because it has a lower affinity to lac promoter .

We confirmed that lacIM1 shows weaker repression than LacIWT.(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])
We ligated BBa_K395400 together with Pbad/araC promoter. 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]

Figure4-2-2. Repression efficiency of LacIM1 (BBa_K395401) / LacIWT (BBa_K395402) exposed to arabinose and IPTG. This work is done by Mitsuhiko Odera

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 7: / Line 7: @@
 We confirmed that lacIM1 shows weaker repression than LacIWT.(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])<br>
 We ligated BBa_K395400 together with Pbad/araC promoter.
-Then, we used it together with BBa_I7106(lacI+pL-RBS-GFP) on pSB3K3 (Fig.  3-2-1).
+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.
-Fluorescence was measured, and the result is shown in Fig. 3-2-2.
+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_ver4.png|left|thumb|300px|Figure3-2-1. ]]
+[[Image:Tokyotech_LacIM1_data_ver2.png|center|thumb|300px|Figure4-2-2. Repression efficiency of LacIM1 (BBa_K395401) / LacIWT (BBa_K395402) exposed to arabinose and IPTG.  This work is done by Mitsuhiko Odera ]]
-[[Image:Tokyotech_LacIM1_data_ver2.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 ]]