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971.
Strains of Thermus thermophilus produce unique carotenoids called thermozeaxanthins and their colonies are light-yellow pigmented. Here, we developed a new cloning system allowing for the rapid and convenient detection of recombinants by color screening based on carotenoid production in T. thermophilus. We constructed two cloning vectors that overexpress the crtB gene encoding a phytoene synthase under the strong promoter of the slpA gene. Phytoene synthase is one of essential enzymes for the production of carotenoids. We also isolated a carotenoid-overproducing mutant that formed orange colonies. Because disruption of crtB in the carotenoid-overproducing mutant resulted in white colonies, we used the disruptant as a host strain. Whereas transformants carrying a new cloning vector, pTRK1-PRslpA-crtBcas, grew into unusual red-pigmented colonies probably because of the extreme accumulation of thermozeaxanthins, those carrying the vector with a foreign DNA inserts formed white colonies. Thus, recombinants can be detected easily by color screening (red/white screening) in T. thermophilus. This cloning system requires no additional chromogenic substrate in the medium. We also constructed a promoter-probe vector, pTRK1-crtBmcs-PP, employing the open reading frame of crtB with multiple cloning sites. Using this vector, a series of colony-color phenotypes is observed probably depending on promoter activities of foreign DNA inserts, which enables the rapid probing of promoters. These vectors are useful to simplify cloning procedures and to identify the promoters of different strengths in T. thermophilus. 相似文献
972.
In many animals, germline progenitors are kept undifferentiated to give rise to germline stem cells (GSCs), enabling continuous production of gametes throughout animal life. In the Drosophila ovary, GSCs arise from a subset of primordial germ cells (PGCs) that stay undifferentiated even after gametogenesis has started. How a certain population of PGCs is protected against differentiation, and the significance of its regulatory mechanisms on GSC establishment remain elusive. Here we show that epidermal growth factor receptor (Egfr) signaling in somatic stromal intermingled cells (ICs), activated by its ligand produced in germ cells, controls the size of the PGC pool at the onset of gametogenesis. Egfr signaling in ICs limits the number of cells that express the heparan sulfate proteoglycan Dally, which is required for the movement and stability of the locally-produced stromal morphogen, Decapentaplegic (Dpp, a BMP2/4 homologue). Dpp is received by PGCs and maintains them in an undifferentiated state. Altering Egfr signaling levels changes the size of the PGC pool and affects the number of GSCs established during development. While excess GSC formation is compensated by the adult stage, insufficient GSC formation can lead to adult ovarioles that completely lack GSCs, suggesting that ensuring an absolute size of the PGC pool is crucial for the GSC system. 相似文献
973.
Shinya Nishimura Ki-Ja Hyun Young-Ah Lee Hiromi Tokura 《Biological Rhythm Research》2013,44(3):233-236
A spectral analysis of heart rate was carried out on 11 young female adults in order to evaluate the effects of bright light exposure on autonomic nervous activity. Bright light (5,000 lx) was provided by fluorescent lamps during the daytime (07:00–15:00) on day 1. Dim light (200 lx) was given on day 2. High frequency components (HF: 0.15–0.4Hz) were used as a marker of parasympathetic activity and the ratio of low frequency (LF: 0.04–0.15 HZ) to high frequency (LF/HF) as an indicator of sympathetic activity. The average value during the sleep period (23:30–06:30) was compared following diurnal exposure to bright or dim light. HF component was significantly greater from 23:30 to 02:00 after diurnal exposure of bright light, being accompanied by lower heart rate during these periods. There existed negative correlation between heart rate and HF component from 23:30 to 02:00 under diurnal exposure to bright and dim lights. The results indicate that bright light exposure during the daytime (07:00–15:00) could enhance parasympathetic activity around midnight. 相似文献
974.
Kozo Ohtsuki Makoto Kawabata Kuniko Taguchi Hiroshi Kokura Shinya Kawamura 《Bioscience, biotechnology, and biochemistry》2013,77(10):2471-2475
S-Methylmethionine (MMS, an anti-ulcer factor, Vitamin u) was determined in the extracts of various kinds of teas, such as green teas, black teas and oolong teas, using an amino acid analyzer for physiological-fluid analysis or for rapid analysis. MMS in the column eluates was confirmed to be dimethyl sulfide by a gas-chromatographic method with a flame photometric detector. The quantity of MMS obtained from the various green teas depended on the quality and the freshness, i.e.fresh, high-quality gyokuro, 15.7 to 24.5 mg%; fresh sen-cha, 7.0 to 10.3 mg%; and the other green teas 1 to 6mg%. Oolong tea and black tea did not contain MMS.The extraction conditions for and the heat-stability of MMS were also discussed. 相似文献
975.
Shinya Takanao Syouta Honma Takuma Miura Chisana Ogawa Hayuki Sugimoto Kazushi Suzuki 《Bioscience, biotechnology, and biochemistry》2013,77(3):524-532
In order to elucidate the roles of ChiP, ChiQ, and ChiX in chitin utilization by Serratia marcescens 2170, the construction of single-gene deletion mutants of the chiP, chiQ, and chiX genes was attempted by allelic exchange mutagenesis. ΔchiP formed smaller clearing zones and ΔchiX formed larger ones than wild-type 2170 on an agar plate containing colloidal chitin. ΔchiP grew slowly on the lower concentration of (GlcNAc)2, and there was essentially no growth on chitin oligosaccharides larger than (GlcNAc)3. The gene product of chiP was detected in the outer membrane fraction, consistently with the hypothesis that chiP encodes outer membrane chitoporin. Deletion of chiQ decreased and that of chiX increased the growth rates on chitin oligosaccharides. These observations strongly suggest that all three genes are involved in chitin utilization and that the deletion mutants obtained in this study might prove useful tools to clarify the details of the chitin utilization system of this bacterium. 相似文献
976.
977.
978.
Shinya Matsuda Naotoshi Yoshiyama Jaana Künnapuu-Vulli Masatsugu Hatakeyama Osamu Shimmi 《Insect biochemistry and molecular biology》2013,43(5):466-473
The pattern of wing venation varies considerably among different groups of insects and has been used as a means of species-specific identification. However, little is known about how wing venation is established and diversified among insects. The decapentaplegic (Dpp)/bone morphogenetic protein (BMP) signaling pathway plays a critical role in wing vein formation during the pupal stages in Drosophila melanogaster. A key mechanism is BMP transport from the longitudinal veins (LVs) to the posterior crossvein (PCV) by the BMP-binding proteins, short gastrulation (Sog) and twisted gastrulation2/crossveinless (Tsg2/Cv). To investigate whether the BMP transport mechanism is utilized to specify insect wing vein patterns in other than Drosophila, we used the sawfly Athalia rosae as a model, which has distinct venation patterns in the fore- and hindwings. Here, we show that Ar-dpp is ubiquitously expressed in both the fore- and hindwings, but is required for localized BMP signaling that reflects distinct wing vein patterns between the fore- and hindwings. By isolating Ar-tsg/cv in the sawfly, we found that Ar-Tsg/Cv is also required for BMP signaling in wing vein formation and retains the ability to transport Dpp. These data suggest that the BMP transport system is widely used to redistribute Dpp to specify wing venation and may be a basal mechanism underlying diversified wing vein patterns among insects. 相似文献
979.
980.
Kimura S Maruyama J Kikuma T Arioka M Kitamoto K 《Biochemical and biophysical research communications》2011,(3):1197-470
Autophagy is a conserved intracellular degradation process of eukaryotic cells. In filamentous fungi, although autophagy has been reported to have multiple physiological roles, it is not clear whether autophagy is involved in the degradation of misfolded proteins. Here, we investigated the role of autophagy in the degradation of misfolded secretory proteins accumulated in endoplasmic reticulum (ER) in the filamentous fungus Aspergillus oryzae. In late-phase cultures, a disulfide bond-deleted mutant of the secretory protein α-amylase AmyB fused with mDsRed that had accumulated in the ER was subsequently delivered to vacuoles, whereas wild-type AmyB-mDsRed was predominantly located at cell walls and septa. To examine the involvement of autophagy in the delivery of mutant AmyB to vacuoles, mutant AmyB-EGFP was expressed in an A. oryzae autophagy-deficient strain (ΔAoatg8). Microscopic examination revealed that the protein delivery to vacuoles did not occur in the absence of autophagic activity, with mutant AmyB-mDsRed forming large spherical structures surrounded by ER membrane. Hence, we conclude that autophagy is responsible for the delivery of misfolded secretory proteins accumulated in the ER to vacuoles for degradation during late-growth phase in A. oryzae. This is the first study to provide evidence that autophagy plays a role in the degradation of misfolded secretory proteins in filamentous fungi. 相似文献