Difference between revisions of "Part:BBa K175005: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_K175005=== | ===Applications of BBa_K175005=== | ||
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| + | ===Characterization of BBa_K175005=== | ||
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| + | A conjugation test was done using the TUDelft iGEM 2009 [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation protocol] to verify that [https://parts.igem.org/wiki/index.php?title=Part:BBa_K175005 K175005] on a pSB4C5 low-copy backbone was transmitted during conjugation. | ||
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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_K175005 K175005] on pSB4C5 (TRI+CAM)<br> | ||
| + | '''Receivers''': DH5 α cells with [https://parts.igem.org/Part:BBa_J23100 J23100] (AMP) <br> | ||
| + | '''Transconjugants A''': DH5 α cells with R751 and [https://parts.igem.org/Part:BBa_J23100 J23100] (TRI+AMP)<br> | ||
| + | '''Transconjugants B''': DH5 α cells with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K175005 K175005] on 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" | ||
| + | |- align="center" | ||
| + | | Set # || Conjugation Efficiency for Conjugation Testing Plasmid 1 || Conjugation Efficiency for wild R751 | ||
| + | |- align="center" | ||
| + | | 1 || 0.00177 || 0.0141 | ||
| + | |- align="center" | ||
| + | | 2 || 0.00224 || 0.0476 | ||
| + | |- align="center" | ||
| + | | 3|| n/a || 0.0356 | ||
| + | |- align="center" | ||
| + | |} | ||
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| + | The reason for the lower efficiency of K175005 on pSB4C5 compared to R751 is the presence of the entry exclusion protein trbK. If a receiver cell gets K175005 on pSB4C5, it can still receive R751 afterward. But if a receiver cell gets R751, it will not accept K175005 on pSB4C5 afterward because trbK expression will block incoming transfers. | ||
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===User Reviews=== | ===User Reviews=== | ||
Revision as of 21:30, 19 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_K175005
Characterization of BBa_K175005
A conjugation test was done using the TUDelft iGEM 2009 [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation protocol] to verify that K175005 on a pSB4C5 low-copy backbone was transmitted during conjugation.
Donors: [http://www.cbs.knaw.nl/databases/nccb/search_bac_plas.aspx NCCBNr# 2350] cells with R751 and K175005 on 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 K175005 on 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 Conjugation Testing Plasmid 1 | Conjugation Efficiency for wild R751 |
| 1 | 0.00177 | 0.0141 |
| 2 | 0.00224 | 0.0476 |
| 3 | n/a | 0.0356 |
The reason for the lower efficiency of K175005 on pSB4C5 compared to R751 is the presence of the entry exclusion protein trbK. If a receiver cell gets K175005 on pSB4C5, it can still receive R751 afterward. But if a receiver cell gets R751, it will not accept K175005 on pSB4C5 afterward because trbK expression will block incoming transfers.
User Reviews
UNIQ3ae01b08fbe8f2e4-partinfo-00000000-QINU UNIQ3ae01b08fbe8f2e4-partinfo-00000001-QINU