Team:UNICAMP-Brazil/Notebooks/October 21

From 2009.igem.org

(Difference between revisions)
(New page: {{:Team:UNICAMP-Brazil/inc_topo}} {{:Team:UNICAMP-Brazil/inc calendar}} __NOTOC__ ==''' YeastGuard '''== ====Yeast experiments: Lysozyme==== * The yeasts of ethanol samples achieved ...)
(Yeast experiments: Methods of analysis)
Line 21: Line 21:
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.
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).
 +
 +
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.

Revision as of 23:14, 21 October 2009

Topo l2.gif topo_r_igem.gif
topo_r_b.gif
Back to Calendar

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).

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