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81.
Gene structure and expression of the MboI restriction--modification system. 总被引:4,自引:1,他引:3 下载免费PDF全文
The genes from Moraxella bovis encoding the MboI restriction--modification system were cloned and expressed in Escherichia coli. Three open reading frames were found in the sequence containing the genes. These genes, which we named mboA, mboB, and mboC, had the same orientation in the genome. Genes mboA and mboC encoded MboI methyltransferases (named M.MboA and M.MboC) with 294 and 273 amino acid residues, respectively. The mboB gene coded for MboI restriction endonuclease (R.MboI) with 280 amino acid residues. Recombinant E.coli-MBOI, which contained the whole MboI system, overproduced R.MboI. R.MboI activity from E.coli-MBOI was 480-fold that of M.bovis. The amino acid sequences deduced from these genes were compared with those of other restriction--modification systems. The protein sequences of the MboI system had 38-49% homology with those of the DpnII system. 相似文献
82.
Effects of in vivo exposure with fenvalerate, esfenvalerate andDDT on hepatic gap junctional intercellular communication (GJIC) in Sprague-Dawley (SD) rats were examined by in vivolin vitro dye-transfer assay and by immunohistochemical staining of connexin 32 (C×32, major liver gap junction protein). Fenvalerate (75 mg/kg/day), esfenvalerate (25 mg/kg/day), DDT (50 mg/kg/day) and corn oil (vehicle control, 5mllkglday) were administered orally once a day. Animals were killed at weeks 1, 2, 4 and 6 after starting the experiment. In the fenvalerate- and esfenvalerate-groups, no compound-related changes in GJIC and C×32 expression were observed. On the contrary, in the DDT-group, average sizes of the dye spread after injection of Lucifer Yellow decreased at weeks 1, 2 and 4, and the area per GJ spot shown by C×32-immunohistochemical staining decreased at weeks 4 and 6. It is concluded that neither fenvalerate nor esfenvalerate inhibits hepatic GJIC with in vivo exposure. 相似文献
83.
84.
Bo Liu Shigeru Nakashima Seiji Ito Yoshinori Nozawa 《Prostaglandins & other lipid mediators》1996,51(4):233-248
Pertussis toxin-insensitive GTP-binding protein was observed to be involved in prostaglandin F2α(PGF2α)-induced phosphoinositide metabolism in Chinese hamster ovary (CHO) cells transfected with PGF2α receptor cDNA (CHO-PGF2α·R cells) (Ito, S. et al. Biochem. Biophys. Res. Commun. 200: 756, 1994). In the present study, we investigated PGF2α-induced PLD activation in CHO-PGF2α·R cells. PLD activation was examined by measuring the production of [3H]phosphatidylbutanol ([3H]PBut), a specific product of the PLD-catalyzed transphosphatidylation reaction. PGF2α-induced [3H]PBut formation was concentration-dependent with the maximal level obtained at 1 μM PGF2α. The maximal [3H]PBut formation was observed at 2 min after addition of PGF2α. Depletion of extracellular Ca2+ with EGTA suppressed PGF2α-induced PLD activation by 50%. PKC inhibitors Ro31–8425 and calphostin C inhibited PGF2α-induced [3H]PBut formation by 50%. PTK inhibitors genistein and herbimycin A failed to inhibit PGF2α-induced PLD activation. A combination of maximal effective concentrations of PGF2α (1 μM) and PMA (100 nM) enhanced PLD activation in an additive manner. Pretreatment of the cells with PMA for 2 h down-regulated PKCα and decreased PGF2α-induced PLD activation. These results suggest that PLD activation by PGF2α is mediated by both PKC-dependent and -independent pathways and that PKCα is involved in the former pathway. 相似文献
85.
T. Ito Naoko Udaka Yoshiaki Inayama Hitoshi Kitamura Masayoshi Kanisawa 《Histochemistry and cell biology》1997,109(1):67-73
We have examined the distribution of calcium-binding proteins (CaBPs) in adult and fetal lungs of Syrian golden hamsters
(Mesocricetus auratus) using immunostaining with confocal laser microscopy and electron microscopy. Single and grouped (neuroepithelial body; NEB)
endocrine cells were distributed from bronchi to alveolar ducts in the adult lung. Serial frozen sections immunostained for
CaBPs in combination with immunostaining for endocrine markers such as calcitonin gene-related peptide, serotonin, PGP9.5,
and synaptophysin revealed that positive immunostaining for calbindin-D28K (CB-D28K) was seen in single endocrine cells and NEBs. However, other so-called EF-hand family CaBPs, parvalbumin and calretinin,
were not detected. Electron microscopically, positive immunoreaction for CB-D28K was mainly in the organelle-free cytoplasmic
matrix of endocrine cells, and partly in nuclei and associated with secretory granules and endoplasmic reticulum. In fetal
developing lungs, endocrine cells appeared first on gestational day 13, and they were positive for all the endocrine markers
used. However, pulmonary endocrine cells were positively immunostained for CB-D28K from gestational days 15 and 16 onward.
In summary, our observations suggest that CB-D28K is a useful marker for endocrine cells of the lung, and CB-D28K could function
as a mediator of endocrine stimulation or calcium homeostasis in pulmonary endocrine cells.
Accepted: 17 June 1997 相似文献
86.
Induction of larval metamorphosis in the sea cucumber Stichopus japonicus by periphitic diatoms 总被引:3,自引:0,他引:3
The mass production of juvenile seeds of the sea cucumber,Stichopus japonicus has recently developed by the SeaFarming Center of Saga Prefecture. Methods for the culture ofperiphicic diatoms have been improved. There are three importantsteps in propagating the diatoms. The first step is theenrichment, with the addition of the nutrient salts, undercontrolled light intensity. The second step is washing withhigh pressure seawater and reversal of the plates. The laststep is elimination of copepods, which feed on diatoms, usinga pesticide. Small periphitic diatoms such as Navicula,Amphora, Achnanthes, and Nitzschia are easily culturedat a density of more than one million cells cm–2, andthese diatoms are able to induce larval metamorphosis andserve as a food source for juvenile sea cucumbers. 相似文献
87.
Izaura Yoshico Hirata Maria Helena Sedenho Cezari Clovis Ryuichi Nakaie Paulo Boschcov Amando Siuiti Ito Maria Aparecida Juliano Luiz Juliano 《Letters in Peptide Science》1995,1(6):299-308
Summary A general procedure, using the commonly employed solid-phase peptide synthesis methodology for obtaining internally quenched fluorogenic peptides with ortho-aminobenzoyl/dinitrophenyl groups as donor-acceptor pairs, is presented. The essential feature of this procedure is the synthesis of an N
-Boc or-Fmoc derivative of glutamic acid with the -carboxyl group bound to N-(2,4-dinitrophenyl)-ethylenediamine (EDDnp), which provides the quencher moiety attached to the C-terminus of the substrate. The fluorescent donor group, ortho-aminobenzoic acid (Abz), is incorporated into the resin-bound peptide in the last coupling cycle. Depending on the resin type used, Abz-peptidyl-Gln-EDDnp or Abz-peptidyl-Glu-EDDnp is obtained. Using the procedure described above, substrates for human renin and tissue kallikreins were synthesised. Spectrofluorimetric measurements of Abz bound to the -amino group of proline showed that strong quenching of Abz fluorescence occurs in the absence of any acceptor group. 相似文献
88.
Yoshiaki Inayama Izumi Tomiyama Hitoshi Kitamura Yukio Nakatani Takaaki Ito Akinori Nozawa Yasuhiro Usuda Masayoshi Kanisawa 《Histochemistry and cell biology》1995,104(3):191-198
The purpose of this study was to investigate alkaline phosphatase (ALPase) reactivity in rabbit airway epithelial cells. Acetone-fixed, methyl benzoate and xylene-cleared (AMeX-treated) paraffin sections of trachea, bronchus, and lung tissue were stained by an azo dye coupling method for ALPase and examined by light microscopy. Electron histochemical staining was also performed in order to study the sensitivity and specificity of reactivity in each cell type. ALPase reactivity at the light microscopic level was observed exclusively in trachco-bronchial basal cells, and not in bronchiolar basal cells. By electron microscopy, ALPase reactivity was noted in 97.9% of basal cells in the trachea, 97.0% of basal cells in the bronchus, and 94.5% of basal cells and 15.4% of Clara cells in the bronchiole. This was also true for dispersed tracheal epithelial cells. Reactivity was rarely observed in ciliated cells, non-goblet-type secretory cells, and undetermined cells. The reactivity was heatlabile, levamisole-sensitive, and of a non-specific type. These findings indicate that basal cells of rabbit trachea and bonchus have fairly high specificity for ALPase of a non-specific isozyme (92.2% and 95.6%, respectively). Therefore, ALPase is considered to be a useful marker for these cells. 相似文献
89.
90.
PI phenotyping by separator isoelectric focusing (SIEF) was performed on a total of 1000 unrelated Japanese individuals from two different areas of Western Japan. The PI M1M4 subtype was observed together with the six common PI M subtypes. PI*M4 was confirmed to be present but rare in the Japanese. Several new PI variants were identified by comparison runs of each variant with previously reported genetic variants. The significance of treatment of serum with dithiothreitol (DTT) followed by iodacetic acid (IAC) in determination of PI variants is also described. 相似文献