Team:LMU-Munich/Inverter

From 2012.igem.org

(Difference between revisions)
 
(7 intermediate revisions not shown)
Line 4: Line 4:
[[Image:LMU-Munich-Invertersign.png|100px|right|link=Team:LMU-Munich/Inverter]]
[[Image:LMU-Munich-Invertersign.png|100px|right|link=Team:LMU-Munich/Inverter]]
 +
 +
Line 13: Line 15:
* To get an explanation of how it works and see the results: [[Team:LMU-Munich/Inverter#Theory and Results|Data and Results]]
* To get an explanation of how it works and see the results: [[Team:LMU-Munich/Inverter#Theory and Results|Data and Results]]
-
* How to compose your individual Inverter, by fusions PCRs and 3a assemblies is explained here: [[Team:LMU-Munich/Inverter#Construct your own Inverter|Construct your own Inverter]]</p>
+
* How to compose your individual Inverter, by fusions PCRs and 3A assemblies is explained here: [[Team:LMU-Munich/Inverter#Construct your own Inverter|Construct your own Inverter]]</p>
 +
 
 +
 
Line 20: Line 24:
<p align="justify">The main component of the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823040 Inverter] is the small RNA RyhB, which natively translationally inhibits the upstream fused region ''uof<sub>CGU</sub>'' of the ''fur'' gene by binding to it and therefore masking Shine Dalgarno sequence. </p>  
<p align="justify">The main component of the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823040 Inverter] is the small RNA RyhB, which natively translationally inhibits the upstream fused region ''uof<sub>CGU</sub>'' of the ''fur'' gene by binding to it and therefore masking Shine Dalgarno sequence. </p>  
 +
{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
Line 29: Line 34:
{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
|style="width: 70%;background-color: #EBFCE4;" |
|style="width: 70%;background-color: #EBFCE4;" |
-
<font color="#000000"; size="2"><p align="justify"> Fig. 1: Organization of fur mRNA and base-pairing between uof and RyhB. The uof and fur reading frames are depicted by white and black arrows, respectively. The sequence comprising nt −110 to +12 is enlarged. The putative SD sequences and start codons of uof and fur are boxed. The -GG- to -GGAGG- mutation in the putative SD of uof is indicated. </p></font>
+
<font color="#000000"; size="2"><p align="justify"> '''Fig. 1: Organization of fur mRNA and base-pairing between uof and RyhB.''' The uof and fur reading frames are depicted by white and black arrows, respectively. The sequence comprising nt −110 to +12 is enlarged. The putative SD sequences and start codons of uof and fur are boxed. The -GG- to -GGAGG- mutation in the putative SD of uof is indicated. </p></font>
<!-- The uof codons UCA6 and AGA7 involved in iron-responsive decoding (see text) and the two consecutive stop codons of uof in the proximal-->
<!-- The uof codons UCA6 and AGA7 involved in iron-responsive decoding (see text) and the two consecutive stop codons of uof in the proximal-->
Line 48: Line 53:
{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
|style="width: 70%;background-color: #EBFCE4;" |
|style="width: 70%;background-color: #EBFCE4;" |
-
<font color="#000000"; size="2"><p align="justify"> Fig. 2:Design and functional principle of the Inverter switch. </p></font>
+
<font color="#000000"; size="2"><p align="justify"> '''Fig. 2: Design and functional principle of the Inverter switch.''' </p></font>
|}
|}
|}
|}
|}
|}
 +
<p align="justify">The β-galactosidase assay below shows the function of this Inverter. 1 mM IPTG is always present, so that the Reporter is fully induced. When grown with increasing amounts of arabinose in the medium, RyhB is produced and inhibits uof<sub>CGU</sub> and consequently the fused reporter in a concentration-dependent manner.</p>
<p align="justify">The β-galactosidase assay below shows the function of this Inverter. 1 mM IPTG is always present, so that the Reporter is fully induced. When grown with increasing amounts of arabinose in the medium, RyhB is produced and inhibits uof<sub>CGU</sub> and consequently the fused reporter in a concentration-dependent manner.</p>
Line 58: Line 64:
<p align="justify">In order to use the Inverter for the promoters and output of choice, it has to be constructed by fusion PCR (see next section).</p>
<p align="justify">In order to use the Inverter for the promoters and output of choice, it has to be constructed by fusion PCR (see next section).</p>
 +
{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
{| style="color:black;" cellpadding="3" width="70%" cellspacing="0" border="0" align="center" style="text-align:left;"
Line 67: Line 74:
{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
{| style="color:black;" cellpadding="0" width="100%" cellspacing="0" border="0" align="center" style="text-align:center;"
|style="width: 70%;background-color: #EBFCE4;" |
|style="width: 70%;background-color: #EBFCE4;" |
-
<font color="#000000"; size="2"><p align="justify"> Fig. 3: β-Galactosidase assay of Inverter with ''lacZα'' as reporter. The higher the arabinose concentration, the higher the translational repression of ''uof<sub>CGU</sub>-lacZα'' </p></font>
+
<font color="#000000"; size="2"><p align="justify"> '''Fig. 3: β-Galactosidase assay of Inverter with ''lacZα'' as reporter.''' The higher the arabinose concentration, the higher the translational repression of ''uof<sub>CGU</sub>-lacZα'' </p></font>
|}
|}
|}
|}
|}
|}
 +
====Construct your own Inverter====
====Construct your own Inverter====

Latest revision as of 14:11, 26 October 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

Team-LMU Photo2.jpg

The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".

IGEM HQ LMU prize.jpg

[ more news ]

Sporenfreunde