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1.
Yutaro Yamagata Yukiko Muramoto Sho Miyamoto Keiko Shindo Masahiro Nakano Takeshi Noda 《Microbiology and immunology》2019,63(5):164-171
Defective interfering (DI) influenza viruses carry a large deletion in a gene segment that interferes with the replication of infectious virus; thus, such viruses have potential for antiviral therapy. However, because DI viruses cannot replicate autonomously without the aid of an infectious helper virus, clonal DI virus stocks that are not contaminated with helper virus have not yet been generated. To overcome this problem, we used reverse genetics to generate a clonal DI virus with a PB2 DI gene, amplified the clonal DI virus using a cell line stably expressing the PB2 protein, and confirmed its ability to interfere with infectious virus replication in vitro. Thus, our approach is suitable for obtaining purely clonal DI viruses, will contribute to the understanding of DI virus interference mechanisms and can be used to develop DI virus‐based antivirals. 相似文献
2.
3.
Preferential binding of E.coli histone-like protein HU alpha to negatively supercoiled DNA. 总被引:3,自引:0,他引:3
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Binding specificity of histone-like HU alpha protein to supercoiled DNA was examined by gel retardation assay and chemical probing with OsO4. The latter method was proved to be a unique means for detecting torsional tension restrained in supercoiled plasmid in the presence of HU alpha. It was shown that HU alpha protein has preferential affinity to negatively supercoiled DNA relative to relaxed, nicked and linearized DNAs. There were two modes for binding of HU alpha to the supercoiled DNA: one was the binding associated with topological changes in DNA and the other was relatively strong binding, probably specific to certain particular structures of DNA. It was suggested that HU in vivo interacts preferentially with the regions deformed under torsional stress or with the metabolically active regions along DNA. 相似文献
4.
In an 18 year old Japanese larch stand, leaf characteristics such as area, weight, gross photosynthetic rate and respiration
rate were studied in order to obtain basic information on estimating canopy photosynthesis and respiration. The leaf growth
courses in area and weight from bud opening were approximated by simple logistic curves. The growth coefficient for the area
growth curve was 0.155–0.175 day−1, while that for the weight growth was 0.112–0.117 day−1. The larger growth coefficient in area growth caused the seasonal change in specific leaf area (SLA) that increased after bud opening to its peak early in May at almost 300 cm2 g−1 and then decreased until it leveled off at about 140 cm2g−1. The change inSLA indicates the possibility that leaf area growth precedes leaf thickness growth. The relationship between the coefficientsa andb of the gross photosynthetic rate (p)-light flux density (1) curve (p=bI/(1+aI)) and the mean relative light flux density (I′/I
0) at each canopy height were approximated by hyperbolic formulae:a=A/(I′/I
0)+B andb=C/(I′/I
0)+D. Leaf respiration rate was also increased with increasingI′/I
0. Seasonal change of gross photosynthetic rate and leaf respiration rate were related to mean air temperature through linear
regression on semilogarithmic co-ordinates. 相似文献
5.
Bacillus subtilis inositol dehydrogenase-encoding gene (idh): sequence and expression in Escherichia coli. 总被引:2,自引:0,他引:2
The Bacillus subtilis inositol dehydrogenase (Idh)-encoding gene (idh) was cloned in the B. subtilis temperate phage, rho 11, and then in Escherichia coli plasmids (pBR322 and pUC118). The nucleotide sequence of the idh gene, which consists of 344 codons and whose product has an Mr of 38,351, was determined. E. coli, bearing pIOL05d15, in which expression of the idh gene is under the control of the lac promoter of pUC118, overproduced an active Idh to approx. 20% of total protein upon addition of isopropyl-beta-D-thiogalactopyranoside. This overproduced enzyme cross-reacted with an anti-Idh antibody, and exhibited the same Mr and substrate specificity as those of the B. subtilis enzyme. 相似文献
6.
M Shimidzu H Shindo U Matsumoto K Mita M Zama 《Archives of biochemistry and biophysics》1985,241(2):692-695
Comparative studies on the conformational stability of histones H1 and H5 have been carried out by monitoring the pH-induced conformational transitions of the proteins by CD and 1H NMR spectroscopies. The transition point of H1 agrees with the pKa of the carboxyl groups of the acidic residues. In contrast, the transition of H5 is associated with the ionization of the histidine residues which exist exclusively in H5, as well as the deionization of the acidic residues. These observations, combined with the result of the deuterium exchange rates of the histidine C-2 protons, led us to conclude that His-25 and His-62, which are buried in the globular domain, play an important role in the conformational stability of histone H5. 相似文献
7.
Distinction and similarity in the structure of histones H1 and H5 as indicated by 13C nuclear-magnetic-resonance spectroscopy 总被引:1,自引:0,他引:1
The 13C nuclear magnetic resonance studies have been carried out on histones H1 and H5, by focusing our interest on possible formation of specific salt bridges between acidic and basic amino acid residues in the proteins and also on the structural difference between the two proteins. The 13C chemical shift and pKa values of the carboxyl group of glutamic acid residues in the histones coincided with those of free glutamic acid. Based on this result and another experiment using completely modified lysine residues in the histones, no evidence for a specific interaction between acidic and basic residues has been found. It has also been shown that the pH-effects of aliphatic and aromatic resonances are quite different between H1 and H5, suggesting that the globular domain of H5 is more stable than that of H1. The correlation time (1.5 ns) for the alpha-carbons of H5 estimated from 13C nuclear Overhauser enhancement was twice as long as that of H1 (0.9 ns), indicating that the backbone in the N-terminal and C-terminal domains of H5 is less mobile than that of H1. 相似文献
8.
Takenobu Ishii Montserrat Ruiz-Torruella Jae Young Kim Hiroyuki Kanzaki Abdullah Albassam Wichaya Wisitrasameewong Satoru Shindo Roodelyne Pierrelus Alireza Heidari Umadevi Kandalam Shin Nakamura Alexandru Movila Dmitriy Minond Toshihisa Kawai 《Journal of cellular and molecular medicine》2023,27(12):1750-1756
Bone remodelling is mediated by orchestrated communication between osteoclasts and osteoblasts which, in part, is regulated by coupling and anti-coupling factors. Amongst formally known anti-coupling factors, Semaphorin 4D (Sema4D), produced by osteoclasts, plays a key role in downmodulating osteoblastogenesis. Sema4D is produced in both membrane-bound and soluble forms; however, the mechanism responsible for producing sSema4D from osteoclasts is unknown. Sema4D, TACE and MT1-MMP are all expressed on the surface of RANKL-primed osteoclast precursors. However, only Sema4D and TACE were colocalized, not Sema4D and MT1-MMP. When TACE and MT1-MMP were either chemically inhibited or suppressed by siRNA, TACE was found to be more engaged in shedding Sema4D. Anti-TACE-mAb inhibited sSema4D release from osteoclast precursors by ~90%. Supernatant collected from osteoclast precursors (OC-sup) suppressed osteoblastogenesis from MC3T3-E1 cells, as measured by alkaline phosphatase activity, but OC-sup harvested from the osteoclast precursors treated with anti-TACE-mAb restored osteoblastogenesis activity in a manner that compensates for diminished sSema4D. Finally, systemic administration of anti-TACE-mAb downregulated the generation of sSema4D in the mouse model of critical-sized bone defect, whereas local injection of recombinant sSema4D to anti-TACE-mAb-treated defect upregulated local osteoblastogenesis. Therefore, a novel pathway is proposed whereby TACE-mediated shedding of Sema4D expressed on the osteoclast precursors generates functionally active sSema4D to suppress osteoblastogenesis. 相似文献
9.
Seasonal changes and yearly gross canopy photosynthetic production were estimated for an 18 year old Japanese larch (Larix leptolepis) forest between 1982 and 1984. A canopy photosynthesis model was applied for the estimation, which took into account the
effect of light interception by the non-photosynthetic organs. Seasonal changes in photosynthetic ability, amount of canopy
leaf area and light environment within the canopy were also taken into account. Amount of leaf area was estimated by the leaf
area growth of a single leaf. The change of light environment within the canopy during the growing season was estimated with
a light penetration model and the leaf increment within the canopy. Canopy respiration and surplus production were calculated
as seasonal and yearly values for the three years studied. Mean yearly estimates of canopy photosynthesis, canopy respiration
and surplus production were 37, 13 and 23 tCO2 ha−1 year−1, respectively. Vertical trend, seasonal changes and yearly values of the estimates were analyzed in relation to environmental
and stand factors. 相似文献
10.