09/05/2014

Hello, dear reader!

Today we’ll get the results of DH5α growth, it contains bio bricks for bioremediation, accumulation and to detect Hg in medium LM (LB + mercury chloride) 2ug/ml, 5ug/ml and 10ug/ml.

Where +, ++ and +++ means low, medium and high growth, respectively. As we could observe, as our biosensor and bioaccumulator don’t hold MerA (mercury reductase) protein MerA, those doesn’t express resistance to higher Hg concentrations than 2ug/ml, unlike our bioremediator, that, as we could evaluate on previous tests, it grows in 10 times higher concentrations. Because of this, our tests for absorption and detection in mass spectrometry and spectrofluorometer will be done in concentrations at lower or equal to 2ug/ml.

Today we transferred transformed DH5α with bio brick BBa_K516030 in liquid and solid medium LB with chloramphenicol 34ug/ml.

In parallel, the test with mercury and the new constructions, we’re also isolating wild bacteria from the contaminated stream and Negro River. Yesterday we made the test with different antibiotics in solid LB, and the result was:

Ampicillin 100ug/ml and streptomycin 20ug/ml:

Tetracycline 12ug/ml:

Chloramphenicol 34ug/ml and kanamycin 50ug/ml:

The result on the following table, comparing the colony’s size in LB medium + antibiotic to the colony’s size on LB medium without antibiotic.

Where +, ++, +++, ++++ and +++++ represents quantitatively the colony similarity between growth on medium with antibiotic related to the colony’s size on medium Luria-Bertani.

We made inoculum in liquid medium of each sample (A1, A2; E1, E2; P1, P2, P3; S1) for total DNA extraction!