Team:UNICAMP-Brazil/Yeastguard/Results

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Lysozyme constitutive expression and Lysozyme expression in response to lactate (DLD promoter) devices were transferred to an yeast expression vector (YEp358 ura+) and then were transformed into the Saccharomyces cerevisiae YF23 ura- strain.
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Lysozyme constitutive expression and Lysozyme expression in response to lactate (DLD promoter) devices were transferred to an yeast expression vector (YEp358 ura+) and then were transformed into the ''Saccharomyces cerevisiae'' YF23 ura- strain.
After being plated on selective medium without uracil, the transformants were selected and transferred to liquid medium also selective.
After being plated on selective medium without uracil, the transformants were selected and transferred to liquid medium also selective.
From the initial OD of ~1.71 the growth of non-induced and lactate-induced yeast was monitored for 5 hours (Figure 1).  
From the initial OD of ~1.71 the growth of non-induced and lactate-induced yeast was monitored for 5 hours (Figure 1).  
[[Image:Graficos.jpg|800px|center]]
[[Image:Graficos.jpg|800px|center]]
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<p style="text-align:justify;font-size:0.9em;">Figure 1:</p>

Revision as of 03:15, 22 October 2009

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The Yeastguard: Results

Confirmed Biobricks


Mechanism of recognition


Parts:

  • BBa_K284002 - JEN1 Promoter from Kluyveromyces lactis -
  • BBa_K284003 - Partial DLD Promoter from Kluyveromyces lactis
Biofusion-digestion.jpg


Devices to JEN1 and DLD promoters characterization:

  • BBa_K284020 - EYFP regulated by lactate responsive promoter (JEN1)
  • BBa_K284021 - EYFP regulated by lactate responsive promoter (DLD)
  • BBa_K284023 - EYFP regulated by constitutive promoter Adh1
20091021 promotores com YFP.jpg


Killing mechanism


Parts:

  • BBa_K284001 - Lysozyme from Gallus gallus
Lysozymedigestionresult.png


Devices to Lysozyme characterization:

  • BBa_K284016 - Lysozyme constitutive expression
  • BBa_K284017 - Lysozyme expression in response to lactate (DLD promoter)
  • BBa_K284015 - Lysozyme expression in response to lactate (JEN1 promoter)
Final adh1lys rwsult.png
20091021 promotores com lis final results.jpg


Yeast experiments: Lysozyme


Lysozyme constitutive expression and Lysozyme expression in response to lactate (DLD promoter) devices were transferred to an yeast expression vector (YEp358 ura+) and then were transformed into the Saccharomyces cerevisiae YF23 ura- strain. After being plated on selective medium without uracil, the transformants were selected and transferred to liquid medium also selective. From the initial OD of ~1.71 the growth of non-induced and lactate-induced yeast was monitored for 5 hours (Figure 1).

Graficos.jpg


Figure 1:


The lysozyme expression were verified by SDS-PAGE (Figure x) and by lactobacilli killing induction (Figure y).


Growth curve



Lactococcus lactis



SDS-PAGE

We made two SDS-PAGE gels, one for the supernatant and the other for the total yeast extract. We couldn't load the pellet ethanol samples maybe because of residual ethanol at the pellets; the alcohol hinder the entry of the sample in the well.

Unfortunately we couldn't see any protein band in the gel neither in the supernatant one nor in the total extract one. Maybe we loaded insufficient amounts of sample.

Geis sds page final result.png



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