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In addition, we performed flow cytometry measurements of cells expressing tester constructs (see Additional file 1 Figure S1A, B).
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To this end we used a tester construct that included genomic SUB coding sequence (see below).
Therefore, reduced proteasomal activity should lead to incomplete proteolysis of a cODC1-containing tester construct.
Using the cyan fluorescent protein (CFP) containing tester construct CFP-cODC1-TDegF-RFP, we observed that pTEV + protease expression led to accumulation of cleavage intermediates in pre1-1 pre1-1 or pre1-1 pre4-1 mutant cells [ 39, 40] indicating incomplete proteolysis of CFP-cODC1-TDeg and F-RFP fragments.
Claw testing was performed over the typical site of sole ulcer (axial sole-bulb junction) and the central part of the dorso-abaxial claw wall of the front limbs by applying a large standard hoof-tester constructed with a force measuring device (KERN FK1K, Kern Kraft & Sohn, Balingen, Germany) welded on to one of the arms.
In order to simulate these conditions a reversed axisymmetrical material tester is designed and constructed.
Indeed it may not be an easy task for the test-maker to observe the right balance between these variables, since striving to control for under-representation may push the tester to add more construct-irrelevance variance to the test, over-representing the ability being measured.
We already constructed a tester plasmid pCETD-12 [ 22].
For tactile sensitivity tests, we constructed an original tester.
We designed and constructed a static tester, in which modularization makes it very convenient to expand function.
A new experimental procedure has been established and a novel Impact Adhesion Tester has been designed and constructed.
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