Difference between revisions of "Part:BBa I714031:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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===Applications of BBa_I714031===
 
===Applications of BBa_I714031===
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===Characterization of BBa_I714031===
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A [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation test] was done by the TUDelft iGEM 2009 team to characterize this part.
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'''Donors''': [http://www.cbs.knaw.nl/databases/nccb/search_bac_plas.aspx NCCBNr# 2350] cells with R751 and [https://parts.igem.org/wiki/index.php?title=Part:BBa_I13522 I13522]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_I714031 I714031]-pSB4C5 (TRI+CAM)<br>
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'''Receivers''': DH5 &alpha; cells with [https://parts.igem.org/Part:BBa_J23100 J23100] (AMP) <br>
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'''Transconjugants A''': DH5 &alpha; cells with R751 and [https://parts.igem.org/Part:BBa_J23100 J23100] (TRI+AMP)<br>
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'''Transconjugants B''': DH5 &alpha; cells with [https://parts.igem.org/wiki/index.php?title=Part:BBa_I13522 I13522]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_I714031 I714031]-pSB4C5 and [https://parts.igem.org/Part:BBa_J23100 J23100] (CAM+AMP)<br>
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The following table shows the calculated conjugation efficiencies (raw data [http://2009.igem.org/Team:TUDelft/19_August_2009#Calin here]):
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{| border="1" align="center"
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|- align="center"
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| Set # || Conjugation Efficiency for [https://parts.igem.org/wiki/index.php?title=Part:BBa_I13522 I13522]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_I714031 I714031]-pSB4C5 || Conjugation Efficiency for wild R751
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|- align="center"
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| 1 || 0.00177 || 0.0141
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|- align="center"
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| 2 || 0.00224 || 0.0476
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|- align="center"
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| 3|| n/a || 0.0356
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|- align="center"
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|}
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The suspected reason for the lower efficiency of [https://parts.igem.org/wiki/index.php?title=Part:BBa_I13522 I13522]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_I714031 I714031]-pSB4C5 compared to R751 is the presence of the entry exclusion protein trbK. If a receiver cell gets [https://parts.igem.org/wiki/index.php?title=Part:BBa_I13522 I13522]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_I714031 I714031]-pSB4C5, it can still receive R751 afterward. But if a receiver cell gets R751, it will not accept [https://parts.igem.org/wiki/index.php?title=Part:BBa_I13522 I13522]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_I714031 I714031]-pSB4C5 afterward because trbK expression will block incoming transfers.
  
 
===User Reviews===
 
===User Reviews===
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*For some data about the efficiency test of conjugation, please see the following link: [http://parts.mit.edu/igem07/index.php/Peking_Hop-Count Data of Conjugation]
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*For some data about the efficiency test of conjugation, please see the following link: [http://2007.igem.org/Peking_Hop-Count Data of Conjugation]

Revision as of 19:25, 21 October 2009

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_I714031

Characterization of BBa_I714031

A [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation test] was done by the TUDelft iGEM 2009 team to characterize this part.

Donors: [http://www.cbs.knaw.nl/databases/nccb/search_bac_plas.aspx NCCBNr# 2350] cells with R751 and I13522+I714031-pSB4C5 (TRI+CAM)
Receivers: DH5 α cells with J23100 (AMP)
Transconjugants A: DH5 α cells with R751 and J23100 (TRI+AMP)
Transconjugants B: DH5 α cells with I13522+I714031-pSB4C5 and J23100 (CAM+AMP)

The following table shows the calculated conjugation efficiencies (raw data [http://2009.igem.org/Team:TUDelft/19_August_2009#Calin here]):

Set # Conjugation Efficiency for I13522+I714031-pSB4C5 Conjugation Efficiency for wild R751
1 0.00177 0.0141
2 0.00224 0.0476
3 n/a 0.0356


The suspected reason for the lower efficiency of I13522+I714031-pSB4C5 compared to R751 is the presence of the entry exclusion protein trbK. If a receiver cell gets I13522+I714031-pSB4C5, it can still receive R751 afterward. But if a receiver cell gets R751, it will not accept I13522+I714031-pSB4C5 afterward because trbK expression will block incoming transfers.

User Reviews

UNIQ03f729bb329f5d60-partinfo-00000000-QINU UNIQ03f729bb329f5d60-partinfo-00000001-QINU

  • For some data about the efficiency test of conjugation, please see the following link: [http://2007.igem.org/Peking_Hop-Count Data of Conjugation]