Team:UNICAMP-Brazil/Notebooks/October 21

YeastGuard

Yeast experiments: Lysozyme

• The yeasts of ethanol samples achieved the optimum OD only today.

• We induced these samples with the same concentrations used in glucose ones, described yesterday.

Yeast experiments: Methods of analysis

Live Lactococcus dead!

• The samples were analyzed by:

I) Aplication of the yeasts culture supernatant in liquid cultures of Lactococcus lactis (the OD of the bacterial culture was monitored for some hours).

II) The glucose samples were also analyzed by inhibition zones formation in solid culture. Unfortunately we have no time to do these tests to the ethanol samples.

III) SDS-PAGE of the supernatant and the total extract of the yeats cells (in case the protein have not been exported) to see if the protein was expressed.

With this tests we expect to see if there is expression of lysozyme under control of the constitutive (Adh1) and the lactate responsive (pDLD) promoters, the last one under lactate induction. The idea of using ethanol as carbon source is to see if there is catabolic repression of the pDLD promoter by glucose, it wouldn't work in glucose samples if this hypotesys were true.

The results of these experiments are documented in the judging form.

Raíssa and Taís