Difference between revisions of "Part:BBa K851004"
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Hawaii 2008 iGEM team [12] attempted before to construct a similar cassette [13] but didn’t succeed. For iGEM UNAM Genomics México 2012 project [14], the Ω Cassette was used in the design of an OR logic gate[15] using a recently described new type of communication system between ''Bacillus Subtilis'' cells called Nanotubes[16].
 
Hawaii 2008 iGEM team [12] attempted before to construct a similar cassette [13] but didn’t succeed. For iGEM UNAM Genomics México 2012 project [14], the Ω Cassette was used in the design of an OR logic gate[15] using a recently described new type of communication system between ''Bacillus Subtilis'' cells called Nanotubes[16].
 +
<br>
 +
<br>
  
  
 +
 +
Second Repetition <br>
 +
<table cellspacing="0" cellpadding="0">
 +
  <tr>
 +
    <td width="209">Spectino    [ng/microliter]</td>
 +
    <td width="83">T0</td>
 +
    <td width="83">T30</td>
 +
    <td width="83">T60</td>
 +
    <td width="83">T90</td>
 +
    <td width="83">T120</td>
 +
    <td width="83">T150</td>
 +
    <td width="83">T180</td>
 +
    <td width="83">T210</td>
 +
    <td width="83">T240</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="right">0</td>
 +
    <td align="right">0.065</td>
 +
    <td align="right">0.12</td>
 +
    <td align="right">0.261</td>
 +
    <td align="right">0.435</td>
 +
    <td align="right">0.57</td>
 +
    <td align="right">0.742</td>
 +
    <td align="right">0.939</td>
 +
    <td align="right">0.966</td>
 +
    <td align="right">1.083</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="right">2.6</td>
 +
    <td align="right">0.045</td>
 +
    <td align="right">0.095</td>
 +
    <td align="right">0.263</td>
 +
    <td align="right">0.408</td>
 +
    <td align="right">0.541</td>
 +
    <td align="right">0.7</td>
 +
    <td align="right">0.897</td>
 +
    <td align="right">0.988</td>
 +
    <td align="right">1.07</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="right">3</td>
 +
    <td align="right">0.048</td>
 +
    <td align="right">0.111</td>
 +
    <td align="right">0.219</td>
 +
    <td align="right">0.428</td>
 +
    <td align="right">0.55</td>
 +
    <td align="right">0.709</td>
 +
    <td align="right">0.851</td>
 +
    <td align="right">1</td>
 +
    <td align="right">1.043</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="right">3.4</td>
 +
    <td align="right">0.048</td>
 +
    <td align="right">0.094</td>
 +
    <td align="right">0.192</td>
 +
    <td align="right">0.371</td>
 +
    <td align="right">0.534</td>
 +
    <td align="right">0.697</td>
 +
    <td align="right">0.844</td>
 +
    <td align="right">0.989</td>
 +
    <td align="right">1.036</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="right">3.6</td>
 +
    <td align="right">0.04</td>
 +
    <td align="right">0.101</td>
 +
    <td align="right">0.214</td>
 +
    <td align="right">0.407</td>
 +
    <td align="right">0.542</td>
 +
    <td align="right">0.679</td>
 +
    <td align="right">0.853</td>
 +
    <td align="right">0.951</td>
 +
    <td align="right">1.067</td>
 +
  </tr>
 +
  <tr>
 +
    <td align="right">3.8</td>
 +
    <td align="right">0.049</td>
 +
    <td align="right">0.102</td>
 +
    <td align="right">0.211</td>
 +
    <td align="right">0.4</td>
 +
    <td align="right">0.552</td>
 +
    <td align="right">0.691</td>
 +
    <td align="right">0.83</td>
 +
    <td align="right">1</td>
 +
    <td align="right">1.006</td>
 +
  </tr>
 +
</table>
 
===REFERENCES===
 
===REFERENCES===
 
<br>
 
<br>

Revision as of 04:31, 27 September 2012


aminoglycoside antibiotic resistance Sm+ Spc+

The Ω Cassette provides resistance to Spectinomycin and Streptomycin; it was obtained from plasmid pHP45Ω [1] as described in [2].


The aminoglycoside antibiotic resistance gene (aadA +)[3,4] of pHP45 was originally carried on a 1.7-kb PvuII-HindIII fragment from the R100.1 plasmid[5]. Plasmid pHP45Ω was constructed in Pierre Prentki and Henry M. Krisch, 1984 [5] as stated in NCBI[6] with this fragment. The addA sequence can be found in [7, 8 ], it is confirmed to be the one contained in pHP45Ω[9] and the complete sequence of pHP45Ω plasmid can be found in [10, 11].

Hawaii 2008 iGEM team [12] attempted before to construct a similar cassette [13] but didn’t succeed. For iGEM UNAM Genomics México 2012 project [14], the Ω Cassette was used in the design of an OR logic gate[15] using a recently described new type of communication system between Bacillus Subtilis cells called Nanotubes[16].


Second Repetition

Spectino [ng/microliter] T0 T30 T60 T90 T120 T150 T180 T210 T240
0 0.065 0.12 0.261 0.435 0.57 0.742 0.939 0.966 1.083
2.6 0.045 0.095 0.263 0.408 0.541 0.7 0.897 0.988 1.07
3 0.048 0.111 0.219 0.428 0.55 0.709 0.851 1 1.043
3.4 0.048 0.094 0.192 0.371 0.534 0.697 0.844 0.989 1.036
3.6 0.04 0.101 0.214 0.407 0.542 0.679 0.853 0.951 1.067
3.8 0.049 0.102 0.211 0.4 0.552 0.691 0.83 1 1.006

REFERENCES


[1] http://www.shigen.nig.ac.jp/ecoli/strain/cvectorDetailAction.do?cvectorId=147
[2] http://2012.igem.org/Team:UNAM_Genomics_Mexico/Notebook/ANDSugar
[3] http://www.ihop-net.org/UniPub/iHOP/bng/174343.html
[4] http://www.wikigenes.org/e/gene/e/1238687.html
[5] Pierre Prentki and Henry M. Krisch (1984). In vitro insertional mutagenesis with a selectable DNA fragment. Gene, 29:303-313
[6] http://www.ncbi.nlm.nih.gov/nuccore/K02163
[7] http://www.uniprot.org/uniprot/P0AG05
[8] http://www.ncbi.nlm.nih.gov/sites/entrez?db=gene&term=1446561
[9] Nancye C. Clark, Ørjan Olsvik, Jana M. Swenson, Carol A. Spiegel and Fred C. Tenover(1999) Detection of a Streptomycin/Spectinomycin Adenylyltransferase Gene ( aadA) in Enterococcus faecalis. Antimicrob. Agents Chemother. 43(1):157.
[10] http://www.ncbi.nlm.nih.gov/nuccore/207855?report=fasta
[11] Pierre Prentki, Anna Bind and Andrée Epstein (1991). Plasmid vectors for selecting ISI-promoted deletions in cloned DNA: sequence analysis of the omega interposon. Gene, 103:17-23.
[12] http://2008.igem.org/Team:Hawaii
[13] http://2008.igem.org/Team:Hawaii/Construction_of_Omega_Interposon_BioBrick
[14] http://2012.igem.org/Team:UNAM_Genomics_Mexico
[15] Link con la página de descripción general del sistema o el de la parte del OR
[16] Dubey GP, Ben-Yehuda S. (2011) Intercellular nanotubes mediate bacterial communication. Cell.;144(4) :590-600

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 5
    Illegal BamHI site found at 2060
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1524
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 499