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Amoebic gill disease in teleost fish is caused by the marine parasite Neoparamoeba perurans. To date, the role of antimicrobial peptides β-defensins and cathelicidins in this infection have not been explored. Using a high-throughput microfluidics quantitative polymerase chain reaction system (Biomark HD™ by Fluidigm), this study aimed to: firstly,...
Of 150 Escherichia coli strains we cultured from specimens taken from cattle in Europe, 3 had elevated MICs against colistin. We assessed all 3 strains for the presence of the plasmid-mediated mcr-1 gene and identified 1 isolate as mcr-1-positive and co-resistant to β-lactam, florfenicol, and fluoroquinolone antimicrobial compounds.
An agarose gel showing the results of a PCR assay designed to detect the mcr-1 resistance encoding gene. Nonsynonymous amino acid substitutions identified in genes associated with resistance to nalidixic acid, fluoroquinolones, and colistin.
I am wondering if anyone here who performs SDS-PAGE has seen this before on their gels post-staining? We make our 12% tricine gels in house, fix them in 25% isopropanol 10% acetic acid, then stained overnight in Coomassie G250 35mM HCl. The gels are then destained in distilled water. We are noticing what seems like "halos" or zones of white around our proteins in the gels. I have images and notes attached regarding the issue. When the peptide is in its neat form, it is in a 1M imidazole, 500mM NaCl, 20mM Tris buffer at pH 8. The peptide has a final concentration of 200mM imidazole when it is in its 1/5 diluted form. We have seen this effect many times before, but are not sure what is it causing it. Is it perhaps due to the presence of imidazole; can the imidazole, or maybe just an overall high salt concentration, cause this effect? We use fresh running buffer, fresh fixative and fresh gel reagents (e.g. new aliquots of APS) for each run. Coomassie is reused and made fresh every month and a half; the Coomassie used here is less than a month old.
Any input or words of wisdom would be greatly appreciated! Many thanks in advance.