Part:BBa_K175001:Experience

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Characterization of BBa_K175001

A conjugation test was done using the TUDelft iGEM 2009 [http://2009.igem.org/Team:TUDelft/ConjugationProtocol conjugation protocol] to verify that expression of trbK blocked conjugation.

Donors: [http://www.cbs.knaw.nl/databases/nccb/search_bac_plas.aspx NCCBNr# 2350] cells with R751 (TRI)
Receivers: DH5 α cells expressing trbK (CAM)
Transconjugants: DH5 α cells with R751 and expressing trbK (TRI+CAM)

Receivers Control: DH5 α cells with I13522 (AMP)
Transconjugants Control: DH5 α cells with R751 and I13522 (TRI+AMP)

The positive control test used the same donors but different receivers, as detailed above.

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

Set #	Conjugation Efficinecy
trbK 1	4.49E-6
trbK 2	2.89E-6
control	0.031

As the graph above shows, expression of trbK reduces the conjugation efficiency by four orders of magnitude. This can be used as receiver side control over conjugation.

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