Difference between revisions of "Part:BBa K175005:Experience"

Latest revision as of 21:32, 19 October 2009

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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-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-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 K175005-pSB4C5	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-pSB4C5 compared to R751 is the presence of the entry exclusion protein trbK. If a receiver cell gets K175005-pSB4C5, it can still receive R751 afterward. But if a receiver cell gets R751, it will not accept K175005-pSB4C5 afterward because trbK expression will block incoming transfers.

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UNIQ2599c3216152fd51-partinfo-00000000-QINU UNIQ2599c3216152fd51-partinfo-00000001-QINU

@@ Line 9: / Line 9: @@
 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.
-'''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>
+'''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]-pSB4C5 (TRI+CAM)<br>
 '''Receivers''': DH5 &alpha; cells with [https://parts.igem.org/Part:BBa_J23100 J23100] (AMP) <br>
 '''Transconjugants A''': DH5 &alpha; cells with R751 and [https://parts.igem.org/Part:BBa_J23100 J23100] (TRI+AMP)<br>
-'''Transconjugants B''': DH5 &alpha; 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>
+'''Transconjugants B''': DH5 &alpha; cells with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K175005 K175005]-pSB4C5 and [https://parts.igem.org/Part:BBa_J23100 J23100] (CAM+AMP)<br>
 The following table shows the calculated conjugation efficiencies (raw data [http://2009.igem.org/Team:TUDelft/19_August_2009#Calin here]):
@@ Line 18: / Line 18: @@
 {| border="1" align="center"
 |- align="center"
-| Set # || Conjugation Efficiency for Conjugation Testing Plasmid 1 || Conjugation Efficiency for wild R751
+| Set # || Conjugation Efficiency for K175005-pSB4C5 || Conjugation Efficiency for wild R751
 |- align="center"
 | 1 || 0.00177 || 0.0141
@@ Line 29: / Line 29: @@
-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.
+The reason for the lower efficiency of K175005-pSB4C5 compared to R751 is the presence of the entry exclusion protein trbK. If a receiver cell gets K175005-pSB4C5, it can still receive R751 afterward. But if a receiver cell gets R751, it will not accept K175005-pSB4C5 afterward because trbK expression will block incoming transfers.
 ===User Reviews===