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11.
Identification of eleven single-strand initiation sequences (ssi) for priming of DNA replication in the F, R6K, R100 and ColE2 plasmids. 总被引:15,自引:0,他引:15
N Nomura H Masai M Inuzuka C Miyazaki E Ohtsubo T Itoh S Sasamoto M Matsui R Ishizaki K Arai 《Gene》1991,108(1):15-22
Based on the ability to complement the poor growth of an M13 phage derivative lacking the complementary strand origin, eleven single-strand initiation sequences (ssi) for DNA replication are identified in the F, R6K, R100 and ColE2 plasmids. Six of them were from F, two from near the gamma and alpha origins (ori) of R6K, two from the vicinity of the basic replicon of R100 and one from near the ori of ColE2. They can be classified into two groups based on the morphology of the plaques and the length of nucleotide (nt) sequences required for ssi activity; one group that gives rise to larger and clearer plaques and can be reduced to nearly 100 nt (seven out of eleven), and another that generates smaller and less clear plaques and requires more than 200 nt for full activity (four out of eleven). Sequence homology is detected among some members from both groups. The possible biological roles of the ssi are discussed. 相似文献
12.
T Masaki H Umehashi H Miyazaki M Takano K Yamakawa S Nakamura 《Japanese journal of medical science & biology》1988,41(1):27-30
A Clostridium perfringens-like strain was isolated from a case of gas gangrene. The morphological properties and the lecithinase reaction of the isolate were very similar to those of C. perfringens; however, the lecithinase reaction was only slightly suppressed by C. perfringens alpha-antitoxin serum and the organism was identified as Clostridium absonum from its biochemical properties. 相似文献
13.
Oxygen equilibrium study and light absorption spectra of Ni(II)-Fe(II) hybrid hemoglobins 总被引:1,自引:0,他引:1
Ni(II)-Fe(II) hybrid hemoglobins, in which hemes in either the alpha or beta subunit are substituted with Ni(II) protoporphyrin IX, have been prepared and characterized. Since Ni(II) protoporphyrin IX binds neither oxygen nor carbon monoxide, the oxygen equilibrium properties of the Fe subunit in these hybrid hemoglobins were specifically determined. K1 values, namely the equilibrium constants for the first oxygen molecule to bind to hemoglobin, agreed well for these hybrid hemoglobins with the K1 value of native hemoglobin A in various conditions. Therefore, Ni(II) protoporphyrin IX in these hybrid hemoglobins behaves like a permanently deoxygenated heme. Both Ne-Fe hybrid hemoglobins bound oxygen non-co-operatively at low pH values. When the pH was raised, alpha 2 (Fe) beta 2 (Ni) showed co-operativity, but the complementary hybrid, alpha 2 (Ni) beta 2 (Fe), did not show co-operativity even at pH 8.5. The light absorption spectra of Ni(II)-Fe(II) hybrid hemoglobins indicated that the coordination states of Ni(II) protoporphyrin IX in the alpha subunits responded to the structure of the hybrid, whereas those in the beta subunits were hardly changed. In a deoxy-like structure (the structure that looks like that observed in deoxyhemoglobin), four-co-ordinated Ni(II) protoporphyrin IX was dominant in the alpha (Ni) subunits, while under the conditions that stabilized an oxy-like structure (the structure that looks like that observed in oxyhemoglobin), five-co-ordinated Ni(II) protoporphyrin IX increased. The small change observed in the absorption spectrum of the beta (Ni) subunits is not related to the change of the co-ordination number of Ni(II) protoporphyrin IX. Non-co-operative binding of oxygen to the beta subunits in alpha 2 (Ni) beta 2 (Fe) accompanied the change of absorption spectrum in the alpha (Ni) subunits. We propose a possible interpretation of this unique feature. 相似文献
14.
15.
Cloning in Saccharomyces cerevisiae of a cycloheximide resistance gene from the Candida maltosa genome which modifies ribosomes. 总被引:1,自引:1,他引:0 下载免费PDF全文
We have previously shown that cycloheximide resistance can be induced in a strain of Candida maltosa by modifying ribosomes (M. Takagi, S. Kawai, Y. Takata, N. Tanaka, M. Sunairi, M. Miyazaki, and K. Yano, J. Gen. Appl. Microbiol. 31:267-275, 1985). The present paper describes the cloning of the gene involved in this resistance (designated RIM-C for ribosome modification by cycloheximide) by using a host-vector system of Saccharomyces cerevisiae. 相似文献
16.
After a large amount of aldosterone was injected into a male rabbit, urine was collected for 48 h. Separation of urinary aldosterone metabolites into monoglucosiduronate fraction and monosulphate fraction was carried out by a combination of countercurrent distribution and DEAE-Sephadex A-25 column chromatography. Each fraction was hydrolyzed with enzyme and free steroids released were separated by Sephadex LH-20 column chromatography. The free steroid was then identified by gas chromatography-mass spectrometry. In monoglucosiduronate fraction, 3 alpha, 5 beta-tetrahydroaldosterone and 3 beta, 5 alpha-tetrahydroaldosterone were found. On the other hand, 3 alpha, 5 beta-tetrahydroaldosterone was the only aglycone detected in monosulphate fraction. These findings comfirmed results in the preceding paper, where the free steroid was characterized on the basis of the mobility of the steroid and its derivatives on paper chromatography. 相似文献
17.
In the presence of phenobarbital (PB) at 3 mM, hepatocytes isolated from adult rats by a collagenase-perfusion technique survived well on plastic dishes for at least 49 days after initiation of primary culture. PB at concentrations less than 3 mM was ineffective for the maintenance of hepatocytes, and the maintenance of them was attained only in the continuous presence of 3 mM PB. The hepatocytes surviving in the presence of 3 mM PB were morphologically indistinguishable from the hepatocytes after 1-day attachment period, except for the presence of prominent nucleoli in the former. Although both the albumin secretion and tyrosine aminotransferase (TAT) activities of the cells decreased gradually up to day 7 with time in culture, both were thereafter maintained at relatively high levels at least up to day 35 of primary culture. The addition of 10 microM dexamethasone caused a 3-5-fold induction in TAT activity, and the cells were capable of responding to the hormone in this manner at least up to day 28 of primary culture. Furthermore, the cells also had glucose-6-phosphatase activity, even though the level of this enzyme activity was relatively low as compared with that of TAT activity. Survival of hepatocytes in the presence of 3 mM PB was further enhanced by simultaneous addition of dexamethasone (10 microM) and insulin (10 micrograms/ml). The sensitivity of hepatocytes to 3'-methyl-4-dimethylaminoazobenzene (0.24 mM) was remarkably reduced by treatment with PB at 3 mM. PB treatment decreased efficiently the falling rate of total cytochrome P-450 content, but did not induce P-450PB, which is the specific form of cytochrome P-450 induced by PB, in primary cultured hepatocytes. On the other hand, 3-methylcholanthrene (MC, 10 microM) caused an increase of both contents of total cytochrome P-450 and P-450MC, which is the specific form of cytochrome P-450 induced by MC, in primary cultured hepatocytes. However, MC was ineffective for the maintenance of hepatocytes in primary culture. The possible biological actions of PB on primary cultured hepatocytes are discussed on the basis of the experimental data obtained. 相似文献
18.
Pyroglutamyl-lysyl-leucyl-argininal (Pyr-Lys-Leu-Argal) immobilized on gel matrix through the epsilon-amino group of its lysine residue was shown to be an efficient biospecific affinity adsorbent for purification of urokinase. Pyr-Lys-Leu-Argal dibutylacetal, a precursor of this immobilized ligand, was synthesized by a fragment condensation procedure, in which one of the thermolysin-digestion products of leupeptin dibutylacetal, H-Leu-Argal dibutylacetal, was used as a key intermediate. The precursor was coupled to CH-Sepharose 4B with the aid of a water-soluble carbodiimide, and its acetal protecting group was then removed by mild acid treatment to free the essential aldehyde function. The Sepharose derivative thus prepared was shown to adsorb urokinase selectively and effectively from a crude human urine preparation at neutral pH and to release the bound enzyme under mild acidic conditions. The present technique afforded a highly purified urokinase preparation abundant in the high-molecular form with 90% recovery. The complex formed between urokinase and the immobilized ligand was found to have a dissociation constant of about 2 X 10(-4)M. 相似文献
19.
Kaoru Miyazaki Keisuke Mashima Nobuhiko Yamashita Jinpei Yamashita Takekazu Horio 《In vitro cellular & developmental biology. Plant》1985,21(1):62-66
Summary We have previously reported the transformation by Rous sarcoma virus of a cloned epithelial cell line (BRL) established from
Buffalo rat liver by H. Coon. The nontransformed (BRL) and transformed (RSV-BRL) cells grew at comparable rates in culture,
whereas only the transformed cells were tumorigenic in vivo. We report here on the existence in rat and mouse sera of a growth
inhibitor for the nontransformed BRL cells. The transformed BRL cells (RSV-BRL) were insensitive to this inhibitor. The inhibitory
activity was not prominent in sera from other species of animals tested except for rabbit; this serum inhibited the growth
of RSV-BRL cells more strongly than that of BRL cells. The growth inhibitor was partially purified from rat serum. It is a
protein free of lipid and has a molecular weight of about 220 000. The inhibitor could be separated into three components
of pI 4.6, 5.2 (major) and 5.6 by isoelectric electrophoresis.
EDITOR'S STATEMENT Although compelling theoretical arguments sometimes can be made for the likely existence of growth-inhibitory
substances of physical relevance in the control of cell proliferation, experiments aimed at identifying and studying such
factors often are difficult to design and interpret, and little strong data exists to suggest that growth-inhibitory substances
are important regulatorsin vivo. The information presented in this paper represents a start toward developing a useful system for studying growth-inhibitory
factor. David W. Barnes 相似文献
20.
Structure and antitumor activity of an alkali-soluble polysaccharide from Cordyceps ophioglossoides 总被引:3,自引:0,他引:3
H Yamada N Kawaguchi T Ohmori Y Takeshita S Taneya T Miyazaki 《Carbohydrate research》1984,125(1):107-115
A water-insoluble extracellular glucan (CO-1) was isolated from the precipitate formed on incubation of the culture filtrate of Cordyceps ophioglossoides at 37 degrees for 19 h. CO-1 was homogeneous as judged by h.p.l.c., electrophoresis, and ultracentrifugation, and the average molecular weight was determined by h.p.l.c. to be 632,000. The 1H- and 13C-n.m.r. and the i.r. spectra indicated that the glucosidic linkages in CO-1 were beta. From the results of methylation analysis, Smith degradation, n.m.r. studies, and enzymic hydrolysis, it was concluded that CO-1 is composed of a backbone of (1----3)-linked beta-D-glucopyranosyl residues with a beta-D-glucopyranosyl group attached to O-6 of every second D-glucopyranosyl residue of the backbone. CO-1 strongly inhibited the growth of Sarcoma 180 solid-type tumor. CO-1 polyalcohol, which was prepared by Smith degradation of CO-1, exhibited more-potent antitumor activity than CO-1. The absorption maximum of Congo Red shifted significantly in the presence of CO-1. 相似文献