We extracted dsTrxz/hpTrxz produced by M-JM109-LacY containing BBa_K2377004 (Fig.1)/BBa_K2377005 (Fig.2). M-JM109-LacY and M-JM109-LacY containing pDP (Fig.3) are control groups.
Products are shown in Fig.4. DsTrxz and hpTrxz are approximately 500bp. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (Yellow box)
Fig.4 Gel electrophoresis picture of dsRNA extraction products. (1) BBa_K2377004: DPTrxz, mini-preparation. (2) BBa_K2377005: TrxzL, mini-preparation. (3) BBa_K2377004: DPTrxz, mass isolation. (4) BBa_K2377005: TrxzL, mass isolation. (5) BBa_K2377016: pDP mini-preparation. (6) M-JM109-LacY, mini-preparation. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (yellow box)
To verify the function of dsTrxz, we did the plant treatment experiment.
+
</p>
+
<p class="mdc-typography--body2">
+
Three groups of Arabidopsis are shown below:
+
</p>
+
<figure>
+
<table>
+
<thead>
+
<tr>
+
<th>group/treatment</th>
+
<th>Days in MS media</th>
+
<th>Days in MS + dsTrxz</th>
+
</tr>
+
</thead>
+
<tbody>
+
<tr>
+
<th>group 1</th>
+
<td>7</td>
+
<td>5</td>
+
</tr>
+
<tr>
+
<th>group 2</th>
+
<td>14</td>
+
<td>21</td>
+
</tr>
+
<tr>
+
<th>group 3</th>
+
<td>22</td>
+
<td>30</td>
+
</tr>
+
</tbody>
+
</table>
+
<figcaption>Data of group 2 is perfect but lost. Here we show the data of group 1 and group 3.</figcaption>
+
</figure>
+
<h2 class="mdc-typography--title">Group 1</h2>
+
<p class="mdc-typography--body2">
+
We seperated group 1 into six subgroups. Each group is treated with different concentration of dsTrxz:
+
</p>
+
<figure>
+
<table>
+
<thead>
+
<tr>
+
<th>Subgroup</th>
+
<th>Concentration (ng/L)</th>
+
</tr>
+
</thead>
+
<tbody>
+
<tr>
+
<th>1</th>
+
<td>0</td>
+
</tr>
+
<tr>
+
<th>2</th>
+
<td>0.1</td>
+
</tr>
+
<tr>
+
<th>3</th>
+
<td>1</td>
+
</tr>
+
<tr>
+
<th>4</th>
+
<td>10</td>
+
</tr>
+
<tr>
+
<th>5</th>
+
<td>100</td>
+
</tr>
+
<tr>
+
<th>6</th>
+
<td>1000</td>
+
</tr>
+
</tbody>
+
</table>
+
</figure>
+
<p class="mdc-typography--body2">
+
We did this experiment to optimize concentration of dsTrxz treatment. As we can see in figure 5, LWRs with dsTrxz treatment are higher than those without dsTrxz treatment. LWRs are relative stable with consentration higher than 1ng/L.
Fig.6 Comparison of control group and treatment group. 2a. Treatment group shows significant poorer growth vigor than control group. 2b. Leaves turn yellow one month after dsTrxz treatment.
+
</figcaption>
+
</figure>
+
<p class="mdc-typography--body2">
+
We measured number of leaves (Fig.7a), average length of longest leaves (Fig. 7b) and average LWR (Fig. 7c) of contol group and dsTrxz group. Results show significant differences after dsTrxz treatment.
Fig.7 Arabidopsis 1 month after growing on media with 10ng/L dsTrxz. (a) Number of leaves shows changes of growth vigor after dsTrxz treatment. (b) Average length of longest leaves shows changes of growth vigor agter dsTrxztreatment. (c) Average LWR shows significant whiting phenotype after dsTrxz treatment.
+
</figcaption>
+
</figure>
+
</article>
+
</main>
</html>
</html>
{{Lanzhou/Footer}}
{{Lanzhou/Footer}}
Revision as of 16:10, 1 November 2017
Lanzhou
Lanzhou2017
DsRNA Extraction
We extracted dsTrxz/hpTrxz produced by M-JM109-LacY containing BBa_K2377004 (Fig.1)/BBa_K2377005 (Fig.2). M-JM109-LacY and M-JM109-LacY containing pDP (Fig.3) are control groups.
Products are shown in Fig.4. DsTrxz and hpTrxz are approximately 500bp. We can see in the picture that dsTrxz and hpTrxz are successfully extracted. (Yellow box)
Plant Treatment
To verify the function of dsTrxz, we did the plant treatment experiment.
Three groups of Arabidopsis are shown below:
group/treatment
Days in MS media
Days in MS + dsTrxz
group 1
7
5
group 2
14
21
group 3
22
30
Data of group 2 is perfect but lost. Here we show the data of group 1 and group 3.
Group 1
We seperated group 1 into six subgroups. Each group is treated with different concentration of dsTrxz:
Subgroup
Concentration (ng/L)
1
0
2
0.1
3
1
4
10
5
100
6
1000
We did this experiment to optimize concentration of dsTrxz treatment. As we can see in figure 5, LWRs with dsTrxz treatment are higher than those without dsTrxz treatment. LWRs are relative stable with consentration higher than 1ng/L.
Group 2
The picture (Fig. 6) of Control group and dsTrxz (+) group shows significant differeces caused by dsTrxz treatment (10ng/L).
We measured number of leaves (Fig.7a), average length of longest leaves (Fig. 7b) and average LWR (Fig. 7c) of contol group and dsTrxz group. Results show significant differences after dsTrxz treatment.