Difference between revisions of "Part:BBa K1351019"
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<partinfo>BBa_K1351019 short</partinfo> | <partinfo>BBa_K1351019 short</partinfo> | ||
− | Artificial constructed complementary RNA which should bind the mRNA of the SdpI immunity protein [https://parts.igem.org/Part:BBa_K1351017 (BBa_K1351017)] in ''Bacillus subtilis'' in order to disturb the | + | Artificial constructed complementary RNA which should bind the mRNA of the SdpI immunity protein [https://parts.igem.org/Part:BBa_K1351017 (BBa_K1351017)] in ''Bacillus subtilis'' in order to disturb the translation of immunity. Expected was a loss of immunity in ''B. subtilis'' against its own cannibalism toxin SDP [https://parts.igem.org/Part:BBa_K1351043 (BBa_K1351043)] over time. |
− | + | ||
− | + | ||
− | < | + | This part was generated with the RFC10 standard and has the following prefix and suffix: |
+ | |||
+ | {| | ||
+ | |prefix with EcoRI, NotI and XbaI: | ||
+ | |<span style="color:blue">GAATTC</span><span style="color:green">GCGGCCGC</span>T<span style="color:red">TCTAGA</span>G | ||
+ | |- | ||
+ | |suffix with SpeI, NotI and PstI: | ||
+ | |T<span style="color:red">ACTAGT</span>A<span style="color:green">GCGGCCG</span><span style="color:blue">CTGCAG</span> | ||
+ | |} | ||
+ | Sites of restriction enzymes generating compatible overhangs have the same color: <span style="color:blue">EcoRI</span> and <span style="color:blue">PstI</span> in blue, <span style="color:green">NotI</span> in green, <span style="color:red">XbaI</span> and <span style="color:red">SpeI</span> in red. | ||
+ | |||
+ | This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus]. | ||
+ | |||
Results: [http://2014.igem.org/Team:LMU-Munich/Results LMU-2014-Results] | Results: [http://2014.igem.org/Team:LMU-Munich/Results LMU-2014-Results] | ||
+ | |||
+ | First results of this BioBrick are depicted in the following graph: | ||
+ | |||
+ | [[File:Sdpianti.JPG|600px]] | ||
+ | |||
+ | The graph shows growth curves of ''Bacillus subtilis'' wildtype, a ''SdpI'' deletion strain, a strain with our BioBrick [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1351019 BBa_K1351019] as well as a strain carrying our BioBrick [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1351017 BBa_K1351017]. | ||
+ | The results were performed with our protocoll: [https://static.igem.org/mediawiki/2014/c/c7/LMU-Munich14_Luminescence_Assay.pdf Lumi Assay] in MCSE and consist of three replicas of three measurements. The induction of was performed with 0.2% xylose. | ||
+ | These results show that our BioBrick [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1351019 BBa_K1351019] has no impact on the ''SdpI'' immunity. Expected was a decrease of immunity in late stationary phase comparible with the deletion of ''SdpI''. | ||
+ | <br><br> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== |
Latest revision as of 17:56, 30 October 2014
Reverse complementary RNA sequence which binds the mRNA of the SdpI immunity
Artificial constructed complementary RNA which should bind the mRNA of the SdpI immunity protein (BBa_K1351017) in Bacillus subtilis in order to disturb the translation of immunity. Expected was a loss of immunity in B. subtilis against its own cannibalism toxin SDP (BBa_K1351043) over time.
This part was generated with the RFC10 standard and has the following prefix and suffix:
prefix with EcoRI, NotI and XbaI: | GAATTCGCGGCCGCTTCTAGAG |
suffix with SpeI, NotI and PstI: | TACTAGTAGCGGCCGCTGCAG |
Sites of restriction enzymes generating compatible overhangs have the same color: EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red.
This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus].
Results: [http://2014.igem.org/Team:LMU-Munich/Results LMU-2014-Results]
First results of this BioBrick are depicted in the following graph:
The graph shows growth curves of Bacillus subtilis wildtype, a SdpI deletion strain, a strain with our BioBrick BBa_K1351019 as well as a strain carrying our BioBrick BBa_K1351017.
The results were performed with our protocoll: Lumi Assay in MCSE and consist of three replicas of three measurements. The induction of was performed with 0.2% xylose.
These results show that our BioBrick BBa_K1351019 has no impact on the SdpI immunity. Expected was a decrease of immunity in late stationary phase comparible with the deletion of SdpI.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]