Part:BBa_K873003:Experience

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Applications of BBa_K873003

　　Because this part contains oriT, the R plasmid nic region is where the relaxosome nicks the plasmid and conjugative transfer by R plasmid machinery begins. The function of this kit (K873003) is: when there is a RP4 plasmid, conjugation will be induced to start. We use BL21 which carry the K873003 plasmid as donor bacteria while DH5 α acts as recipient bacteria, and the helper is the HB101 strain who carry the RP4 plasmid. According to the document, DH5 α anti Nalidixic Acid HB101 has resistance over Ampicillin and Kanamycin, and BL21 has Chloramphenicol resistance. If the conjugation happens, the recipient bacterium will not only anti Nalidixic Acid, but also have Chloramphenicol resistance .

Experimental procedure

Results

　　 　　The TEM image of conjugation:

The agar Luria-Bertani medium of DH5α: -------------------------------------------------------------- The agar Luria-Bertani medium of BL21:

The agar Luria-Bertani medium of HB101: -------------------------------------------------------------- The agar Luria-Bertani medium of conjugation:

We process the picture by the model of our member Haoruo Jia. The results are as follows:

Thus the efficiency of conjugation is: Tranconjugants/BL21 = 0.12347.

It's show that the conjugation system did work.
For more information on how this part operates in our system, please visit [http://2012.igem.org/Team:SEU_A SEU_A ]．

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