Team:UNAM Genomics Mexico/prueba

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<center><h1>'''Nanotubes'''</h1></center>  
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<center><h1>'''Bacillus subtilis Results'''</h1></center>  
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<td id="leftcolumn2" align="center"><img src="https://static.igem.org/mediawiki/2012/d/d2/Unamgenomicsnanotubes.jpg" alt="some_text" height="200"/><br /><br /><p>Nanotubes</p></td>
 
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<td  id="contentcolumn2" align="center"><img src="https://static.igem.org/mediawiki/2012/9/96/Unamgenomicsnanotubes1.jpg" alt="some_text" height="200"/><br /><br /><p>Nanotubes</p></td>
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We know that everybody has wetlab problems, just like us. And we actually spend most of the summer looking to standardize the Bacillus subtilis transformation protocol. And we did it! So we are presenting you our new standard:
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[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_competent_cells_protocol | Escherichia coli MC1061 competent cells protocol]]
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Just as people, bacterium need to communicate with others, for this purpose there are many ways, and one of the more recent discovery are Nanotubes that bridge neighboring cells, providing a network for exchange of cellular molecules within and between species. Ben-Yehuda et al discovered these nanotubes in 2011, they show and extraordinary form of communication between Bacillus subtilis. Our team was astonished because of the implications, in the article is described that GFP and calcein, two molecules which cannot leave the cytoplasm, can be transferred to neighboring cells in B. subtillis, it means bacterium can share cytoplasm, the complex network of cells sharing cytoplasm that can be created was our main motivation to create Bacillus Booleanus.<br />
 
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It's still little known about nanotubes, and that's make them a very interesting subject of study, and we are not the first iGEM team interested in working with them, in 2011 Paris_Bettencourt team work with the nanotubes, their goal was to characterize them.<br />
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[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_heat_shock_transformation_protocol Two-step Bacillus subtilis transformation procedure]]
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To accomplish our goal of communicate our logic gates, we contacted [https://medicine.ekmd.huji.ac.il/En/Publications/ResearchersPages/pages/sigalb.aspx Ben Yehuda] group, and [https://2011.igem.org/Team:Paris_Bettencourt Paris Bettencourt iGEM 2011 team], to obtain protocols and work experience.<br />
 
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We work to recreate the formation of nanotubes, but it was impossible to see them because the scanning microscope we have access to, has not enough resolution to see them.<br />
 
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<p class='captionInside'>2011 Gyanendra P. Dubey, Sigal Ben-Yehuda. Intercellular Nanotubes Mediate Bacterial Communication. Cell, 2011; 144 (4): 590 DOI:10.1016/j.cell.2011.01.015<br /></p>
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<p class='captionInside'>2011 Gyanendra P. Dubey, Sigal Ben-Yehuda. Intercellular Nanotubes Mediate Bacterial Communication. Cell, 2011; 144 (4): 590 DOI:10.1016/j.cell.2011.01.015<br /></p>
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We notice that you not only need a plasmid to transform into B. subtilis, but a plasmid that has form multimers in a RecA+ E. coli and should be flanked by AmyE 5' & 3', so that when the plasmid is transformed into Bacillus, it does not get degradated, and it makes an integration into the genome.
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Revision as of 23:04, 14 October 2012


UNAM-Genomics_Mexico


Bacillus subtilis Results





We know that everybody has wetlab problems, just like us. And we actually spend most of the summer looking to standardize the Bacillus subtilis transformation protocol. And we did it! So we are presenting you our new standard:

[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_competent_cells_protocol | Escherichia coli MC1061 competent cells protocol]]




Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_heat_shock_transformation_protocol Two-step Bacillus subtilis transformation procedure




We notice that you not only need a plasmid to transform into B. subtilis, but a plasmid that has form multimers in a RecA+ E. coli and should be flanked by AmyE 5' & 3', so that when the plasmid is transformed into Bacillus, it does not get degradated, and it makes an integration into the genome.

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