Difference between revisions of "Team:HokkaidoU Japan/Proof"
| Line 27: | Line 27: | ||
| − | <br> | + | <br>And also we conducted SDS-PAGE with |
(<a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> + <a href="http://parts.igem.org/Part:BBa_K2015008">BBa_K2015008</a> / <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> + <a href="http://parts.igem.org/Part:BBa_K2015009">BBa_K2015009</a>) on pSB1C3 of <span style="font-style: italic">E. coli</span> (DH5α). | (<a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> + <a href="http://parts.igem.org/Part:BBa_K2015008">BBa_K2015008</a> / <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> + <a href="http://parts.igem.org/Part:BBa_K2015009">BBa_K2015009</a>) on pSB1C3 of <span style="font-style: italic">E. coli</span> (DH5α). | ||
Let us explain the 3 parts below. <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> part contains lacI expression unit and plac + RBS. The sequences of downstream can be induced strictly by IPTG. | Let us explain the 3 parts below. <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> part contains lacI expression unit and plac + RBS. The sequences of downstream can be induced strictly by IPTG. | ||
| Line 33: | Line 33: | ||
| − | + | <br> | |
| − | < | + | <br> |
| − | <table style="border-style: none"> | + | <br> |
| + | |||
| + | The results are shown the below pictures.The fluorescence of GFPs was not detected any condition | ||
| + | <table style="border-style: none; width:1px; margin-left:auto; margin-right:auto"> | ||
<tr align="center"> | <tr align="center"> | ||
<td style="border-style: none;"> | <td style="border-style: none;"> | ||
<tr> | <tr> | ||
| − | <td style="border-style:none; float:center">< | + | <td style="border-style:none; float:center"><img src="https://static.igem.org/mediawiki/2016/d/da/T--HokkaidoU_Japan--multimerization_BBa_K2015012-BBa_K2015008.png" alt="result1" height="300px" width="auto"></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td style="border-style: none"; align="center"> | + | <td style="border-style: none"; align="center"><span class="small">Fig. 1. Result of IPTG induction<br>BBa_K2015012-BBa_K2015008 on pSB1C3 vector were induced by IPTG and incubated at 37°C. (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG</span></td> |
| − | + | ||
| − | < | + | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | </ | + | |
| − | + | ||
| − | + | ||
| − | </td> | + | |
</tr> | </tr> | ||
| + | </tr> | ||
</table> | </table> | ||
| − | |||
<br clear="all"> | <br clear="all"> | ||
| − | + | ||
| − | + | ||
| − | + | ||
<center> | <center> | ||
<table style="border-style: none"> | <table style="border-style: none"> | ||
| − | <tr align="center"> | + | <tr align="center" style="border-style: none"> |
| − | <td style="border-style: none;"> | + | <td style="border-style: none; "> |
| − | + | <img src="https://static.igem.org/mediawiki/2016/b/be/T--HokkaidoU_Japan--multimerization_BBa_K2015012-BBa_K2015009_25.png" alt="result2" height="300px" width="auto"> | |
| − | <td style="border-style:none; | + | </td> |
| − | </ | + | <td style="border-style: none; "> |
| − | <tr> | + | <img src="https://static.igem.org/mediawiki/2016/f/fa/T--HokkaidoU_Japan--multimerization_BBa_K2015012-BBa_K2015009.png" alt="result3" height="300px" width="auto"> |
| − | + | </td> | |
| + | </tr> | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
</table> | </table> | ||
| + | <span class="small">Fig. 2. BBa_K2015012-BBa_K2015009<br>BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25°C for 24 h (left) <br>and at 37°C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG</span> | ||
</center> | </center> | ||
<br clear="all"> | <br clear="all"> | ||
| − | |||
| − | |||
| − | + | . | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
</span> | </span> | ||
Revision as of 02:23, 20 October 2016
"
Team:HokkaidoU Japan
\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\
We made a functonal unit to regulate gene expression. BBa_K2015012 codes constitutive promoter (BBa_J23101), RBS (BBa_B0032), LacI (BBa_C0012), dT (BBa_B0015), PLac (BBa_R0011) and RBS (BBa_B0034). To insert mRFP at the downstream we identified this construct could regulate the expression (Fig. 1).
And also we conducted SDS-PAGE with (BBa_K2015012 + BBa_K2015008 / BBa_K2015012 + BBa_K2015009) on pSB1C3 of E. coli (DH5α). Let us explain the 3 parts below. BBa_K2015012 part contains lacI expression unit and plac + RBS. The sequences of downstream can be induced strictly by IPTG. BBa_K2015008 consists Self Assembling Regions (SAR, RADA16-I) and mutated GFP. And BBa_K2015009 this part consists Self Assembling Region (SAR, P11-4) and mutated GFP.
The results are shown the below pictures.The fluorescence of GFPs was not detected any condition
Fig. 2. BBa_K2015012-BBa_K2015009
BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25°C for 24 h (left)
and at 37°C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG
.
We made a functonal unit to regulate gene expression. BBa_K2015012 codes constitutive promoter (BBa_J23101), RBS (BBa_B0032), LacI (BBa_C0012), dT (BBa_B0015), PLac (BBa_R0011) and RBS (BBa_B0034). To insert mRFP at the downstream we identified this construct could regulate the expression (Fig. 1).
 |
| Fig. 1. Change of expression by IPTG induction left: not induced, right: induced |
And also we conducted SDS-PAGE with (BBa_K2015012 + BBa_K2015008 / BBa_K2015012 + BBa_K2015009) on pSB1C3 of E. coli (DH5α). Let us explain the 3 parts below. BBa_K2015012 part contains lacI expression unit and plac + RBS. The sequences of downstream can be induced strictly by IPTG. BBa_K2015008 consists Self Assembling Regions (SAR, RADA16-I) and mutated GFP. And BBa_K2015009 this part consists Self Assembling Region (SAR, P11-4) and mutated GFP.
The results are shown the below pictures.The fluorescence of GFPs was not detected any condition
 |
| Fig. 1. Result of IPTG induction BBa_K2015012-BBa_K2015008 on pSB1C3 vector were induced by IPTG and incubated at 37°C. (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG |
|
|
BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25°C for 24 h (left)
and at 37°C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG
.
