Part:BBa_I714031:Experience

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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.

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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]