The Yeastguard: Killing
The killing mechanism was designed based on the contaminants´ features. The lactobacilli are gram-positive bacteria and consequently have specific compounds in the cell wall. In addition, lysozyme is a protein that acts on the cell wall of gram-positive bacteria, catalyzing the hydrolysis of the peptidoglycan. Taking this into account, we decided to use the Lysozyme as a way to kill the contaminants and not cause injury to the yeasts.
The link between the recognition and killing mechanisms is the device that consists of one of the sensitive lactate promoters and the lysozyme (Figure 1).
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Lysozyme from Gallus gallus regulated by JEN1 or DLD promoters from Kluyveromyces lactis
Killing protein: Lysozyme
The lysozyme is going to be amplified by PCR from the mRNA of Gallus gallus. The mRNA should be extracted from the organs that express the protein, so the test will be made with blood, spleen and oviduct. The expected amplicon have approximately 540bp.
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