The confirmation experiment of Conjugation

Experiment Notebook.

The confirmation experiment of Sweet gene

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: Fig1. The experimental procedure of Sweet gene’s confirmation experiment Fig2. The transmission electron microscope image of conjugation Fig3. The TEM image of conjugation The data of Sweet gene’s confirmation experiment version 1.0 Fig4. The agar Luria-Bertani medium of DH5α Fig 5. The agar Luria-Bertani medium of DH5α Fig 6. The agar Luria-Bertani medium of BL21 Fig7. The agar Luria-Bertani medium of BL21 Fig8. The agar Luria-Bertani medium of HB101 Fig9. The agar Luria-Bertani medium of HB101 Fig10. The agar Luria-Bertani medium of conjugation Fig11. The agar Luria-Bertani medium of conjugation We process the picture by the model of our member Haoruo Jia. The results are as follows: Cell Number Total area Accuracy class P1 P2 P3 Level(0-1) DH5α 28 846 Nice 800 100 15 0.6 DH5α 2 162 Nice 800 80 70 0.4 BL21 439 8951 Good 800 80 8 0.32 BL21 765 10421 Acceptable 800 80 3 0.18 HB101 381 8700 Good 800 80 3 0.55 HB101 2255 28262 Good 800 80 3 0.35 Transconjugants 72 648 Good 200 80 5 0.2 Transconjugants 130 1819 Good 1000 80 7 0.2 Thus the efficiency of conjugation is: Tranconjugants/BL21 = 0.12347.