Citations

Abstract

Perhaps half of the proteins encoded by the human genome are phosphorylated, and this post-translational modification helps regulate vital processes such as signal transduction and the cell cycle. However, monitoring phosphorylation by Western blotting can be frustrating. Although anti-phosphotyrosine antibodies are reliable, antibodies against phosphoserine and phosphothreonine tend to be sequence dependent. A number of non-antibody-based dyes have been introduced, but high backgrounds and poor sensitivities limit their popularity. 32 P-labeling is a context-independent alternative to Westerns, but its toxicity can perturb cells, and few relish the hassle of working with this beta emitter. Another option, mass spectrometry (MS), suffers from low ionization and fragmentation of phosphopeptides, necessitating sophisticated instrumentation not usually available for routine use. Undaunted by this litany of disappointment in phosphoprotein detection, Iliuk et al. describe sensitive phosphorylation analysis via a modified Western blotting method. The technique, published in Molecular and Cellular Proteomics, uses a soluble hyperbranched nanopolymer called pIMAGO. The reagent is functionalized with titanium ions, which have affinity and selectivity for phosphate groups. The authors previously used this property for enriching phosphopeptides before MS, and

Seasonal cycle of inbreeding and recombination of the parasitic mite Varroa destructor in honeybee colonies and its implications for the selection of acaricide resistance

“…5 μl of proteinase K was then added in each sample. Total mite DNA was isolated from the individuals using standard Chelex thermocycling conditions (Walsh et al, 1991). Four novel polymorphic microsatellite markers (Vdes01-04, Table 2) were designed from the published genome of V.…”

Intra-Colonial Viral Infections in Western Honey Bees (Apis Mellifera)

“…The phenotyped individuals were dissected directly upon sampling. DNA from 486 collected pupae (purple eye stage) was isolated using a hind leg per individual according to the Chelex extraction protocol described by Walsh et al [ 33 ]. The patrilines of the pupae were reconstructed using microsatellite markers, and the relationship between mite infestation traits and brood genotypes was tested.…”

Erysiphe deutziicola sp. nov. (Erysiphaceae, Ascomycota), a powdery mildew species found on Deutzia parviflora (Hydrangeaceae) with unusual appendages

“…Genome DNA was extracted using chasmothecia of HMJAU-PM91860 and conidia and mycelia from the asexual specimen HMJAU-PM91861 by the Chelex-100 method (Walsh et al 1991; Hirata and Takamatsu 1996). Two specimens of Erysiphedeutziae on Deutziaparvifloravar.amurensis (Nguyen et al 2018) were also used for the DNA extraction and amplification, since 28S rDNA sequences of E.deutziae were not yet available in GenBank.…”