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Takahashi H.,Hiroshima University | Takahashi H.,Japan Science and Technology Agency | Takahashi H.,Japan National Food Research Institute | Satoh T.,Koken Ltd | And 10 more authors.
BioTechniques | Year: 2016

Prevention of airborne contamination has become an important factor in biotechnology; however, conventional laminar-airflow cabinets (LAF-cabinets) are no longer sufficient as a countermeasure against nano-sized airborne contaminants in the laboratory. Here we present a bench-top extra-cleanroom classified as ISO-1 that can prevent contamination from airborne nanoparticles. This bench-top extra-cleanroom consists of a novel clean-zone-creating system that is equipped with nanofibrous, nonwoven filters. In addition, the cleanroom is also equipped with an ionizer to prevent plasticware from collecting dust by electrostatic charge attraction. This combination of features allows the cleanroom to prevent DNA contamination derived from airborne nanoparticles. Our extra-cleanroom with ionizer could be useful in various areas of biotechnology that are easily affected by airborne contaminants. © 2016, Eaton Publishing Company. All rights reserved. Source


Sakumoto R.,National Institute of Agrobiological SciencesIbaraki | Hayashi K.-G.,National Institute of Agrobiological SciencesIbaraki | Saito S.,National Institute of Agrobiological SciencesIbaraki | Kanahara H.,National Institute of Agrobiological SciencesIbaraki | And 2 more authors.
Journal of Reproduction and Development | Year: 2015

Heat stress compromises fertility during summer in dairy and beef cows by causing nutritional, physiological and reproductive damages. To examine the difference in endometrial conditions in cows between summer and autumn, gene expression profiles were compared using a 15 K bovine oligo DNA microarray. The trial was conducted in the summer (early in September) and autumn (mid-November) seasons of two consecutive years (2013–2014) in Morioka, Japan. Endometrial samples were collected from the cows using a biopsy technique. The expressions of 268 genes were significantly higher in the endometrium collected in summer than those collected in autumn, whereas the expressions of 369 genes were lower (P<0.05 or lower). Messenger RNA expressions of glycoprotein 2 (GP2), neurotensin (NTS),E-cadherin (CDH1) and heat shock 105kDa/110kDa protein 1 (HSPH1) were validated by quantitative real-time PCR. Transcripts of GP2 and NTS were more abundant in the endometrium from summer than in the endometrium from autumn (P < 0.05). In contrast, the mRNA expressions of CDH1 were lower (P < 0.05) and those of HSPH1 tended to be low (P = 0.09) in the endometrium from summer. Immunohistochemical staining showed that GP2, NTS and HSPH1 were expressed in the endometrial epithelial or glandular epithelial cells. The serum concentrations of NTS collected from the cows in summer were higher than those collected from cows in autumn (P < 0.05). Collectively, the different gene expression profiles may contribute to functional differences in the endometrium between summer and autumn, and the increases in GP2 and NTS may have a relationship with the endometrial deficiency that causes infertility of cows in summer. © 2015 by the Society for Reproduction and Development. Source


Nishizawa-Yokoi A.,National Institute of Agrobiological SciencesIbaraki | Nonaka S.,National Institute of Agrobiological SciencesIbaraki | Nonaka S.,University of Tsukuba | Osakabe K.,National Institute of Agrobiological SciencesIbaraki | And 4 more authors.
Plant Physiology | Year: 2015

Gene targeting (GT) is a useful technology for accurate genome engineering in plants. A reproducible approach based on a positivenegative selection system using hygromycin resistance and the diphtheria toxin A subunit gene as positive and negative selection markers, respectively, is now available. However, to date, this selection system has been applied exclusively in rice (Oryza sativa). To establish a universally applicable positive-negative GT system in plants, we designed a selection system using a combination of neomycin phosphotransferaseII (nptII) and an antisense nptII construct. The concomitant transcription of both sense and antisense nptII suppresses significantly the level of expression of the sense nptII gene, and transgenic calli and plants become sensitive to the antibiotic geneticin. In addition, we were able to utilize the sense nptII gene as a positive selection marker and the antisense nptII construct as a negative selectionmarker for knockout of the endogenous rice genes Waxy and 33-kD globulin through GT, although negative selection with this system is relatively less efficient compared with diphtheria toxin A subunit. The approach developed here, with some additional improvements, could be applied as a universal selection system for the enrichment of GT cells in several plant species. © 2015 American Society of Plant Biologists. All rights reserved. Source

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