Search

Nuclear filament-containing protein 2 (Nuf2p), a centromere-associated protein, is an indispensable component of Ndc80 complex, which is important for stable microtubule–kinetochore attachment, chromosome pairing, and spindle assembly in mitosis. Based upon contig sequences coding for Nuf2p from a pyrosequencing transcriptome of Saccharina japonica, primers were designed for molecular cloning of this kinetochore protein. The open reading frame (ORF) of this gene SjNuf2 was 1,452 bp long, encoding a putative protein consisting of 483 amino acids. Multi-sequence alignment showed SjNuf2p possessed several conserved domains of Nuf2, Spc7, SPT2, and MIT. To illustrate the location of SjNuf2p using immunocytochemistry, its ORF was inserted into a prokaryotic expression vector pET-28a. Then, the recombinant SjNuf2p (rSjNuf2p) was induced to be expressed in Escherichia coli. The rSjNuf2p was verified by Western blotting with the commercial anti-6 × His-tag antibody and then purified using affinity chromatography. The polyclonal antibody of anti-SjNuf2p was raised against the purified rSjNuf2p. Western blotting with this prepared antibody showed the presence of SjNuf2p in the gametophytes of a Saccharina cultivar. SjNuf2p and α/β-tubulin were subsequently co-localized to the paraffin sections of kelp chromosomes with their corresponding antibodies, thus illustrating that SjNuf2p could reside at the kelp centromeres. This study laid a solid foundation for kelp centromere and karyotype analyses.

Keywords: allergen; hypoallergenic varieties; peach; protein quantification; Pru p 3 EN allergen hypoallergenic varieties peach protein quantification Pru p 3 1256 1260 5 05/20/20 20200501 NES 200501 To the Editor, Peach is an important fruit consumed worldwide. In low Pru p 3 content "Hu You 278" (nectarine, 4.02 µg/g) (Figure G1) and peach variety "Xue Bu Dai" (red flesh, 4.00 µg/g, Figure G2), small amounts of Pru p 3 were located in the pericarp layer and less in the mesocarp. The Pru p 3 content in the pulp of a yellow flesh peach cultivar, such as "Jin Shuo", was higher than that in the whole fruit of some nectarines or red flesh peaches. ImmunoCAP results showed that 9/9 were positive to peach, 7/9 positive to Pru p 3 and/or Art v 3, 2/9 positive to Pru p 4 and none of them positive to Pru p 1 (Table). [Extracted from the article]

Objective: To determine Th1/Th2 cytokines and chemokines in patients with allergic diseases and its clinic significance., Methods: Serum levels of IFN-gamma, IL-4, IL-5, IL-13, Eotaxin, RANTES and LTB4 were determined from peripheral blood of 64 allergic patients and 21 healthy controls with ELISA., Results: IL-4, IL-5, IL-13 and Eotaxin, LTB4 were increased significantly in serum of allergic patients compared with those of controls (P<0.05). There were no significant differences in serum levels of IFN-gamma and RANTES between patients and controls (P>0.05). Serum levels of IL-4, IL-5, IL-13 and LTB4 were correlated with each other (P<0.01). Eotaxin, RANTES and IFN-gamma levels were also significantly correlated with each other (P<0.05). LTB4 was correlated with Eotaxin as well (P<0.01)., Conclusion: A wide range of cytokines and chemokines is involved in allergic diseases,which may play their roles in a complex interactive manner.