Part:BBa_K175005:Experience

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.

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.

User Reviews

UNIQcb9ab03c9c7ce9c2-partinfo-00000000-QINU UNIQcb9ab03c9c7ce9c2-partinfo-00000001-QINU