Team:Goettingen/week12-1

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#1 Selection / Swimming - 12th Week

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V07_17


V07_17_1 Analysis of separation assay V07_13_1
  • Experiment:
    From the plates of V07_13_1 were little spots picked. These spots are right at the most outer front of the swimming bacterial halo towards the attractant soaked Whatman paper. The picked colonies were resuspended and plated onto new plates to count the cfus.
  • Observations & Results:
    None of the plates showed the formation of colonies.
V07_17_2: Using an amino acid mix as attractan
  • Experiment:
    According to the swimming assay in methods. M9 agar plates, supplemented with methionine were used. Different strains (∆tar, BL21, and DH10B) were used, containing a variety of plasmids (in case of ∆tar: pSB1C3_rfp and psB1C3_18C_tar; BL21 and DH10B: J61002_18C_rfp and J61002_18C_flhDC). This time, a mix of 18 proteinogenic amino acids was used as chemoattractant.
  • Observations & Results:
    ∆tar: the Tar containing cells showed swimming, whereas the rfp-transformants did not. BL21: flhDC containing E. coli showed clear chemotaxis towards the amino acids, whereas the rfp transformants showed only a little swimming. DH10B: Here, the rfp transformants showed strong swimming and chemotaxis, the flhDC strains did not.

V07_19


Repetition of several swimming assays
  • Experiment
    The strains BL21 and DH10B, containing 18c_rfp and 18c_flhDC constructs, together with ∆tar, containing 18c_rfp and 18c_Tar were dropped onto M9 agar (supplemented with methionine) plates. A variety of chemoattractants was used (tryptone, aspartate, and aa-mix) and soaked into whatman papers. The paper was put in the middle of the plates.


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