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1.
Yusuke Nakamura Michio Ogawa Takahiro Nishide Mitsuru Emi Goro Kosaki Seiichi Himeno Kenichi Matsubara 《Gene》1984,28(2):263-270
The nucleotide sequences of the cloned human salivary and pancreatic α-amylase cDNAs correspond to the continuous mRNA sequences of 1768 and 1566 nucleotides, respectively. These include all of the amino acid coding regions. Salivary cDNA contains 200 bp in the 5′-noncoding region and 32 in the 3′-noncoding region. Pancreatic cDNA contains 3 and 27 bp of 5′- and 3′-noncoding regions, respectively. The nucleotide sequence humology of the two cDNAs is 96% in the coding region, and the predicted amino acid sequences are 94% homologous.Comparison of the sequences of human α-amylase cDNAs with those previously obtained for mouse α-amylase genes (Hagenbuchle et al., 1980; Schibler et al., 1982) showed the possibility of gene conversion between the two genes of human α-amylase. 相似文献
2.
Yukiji Shimojima Hiroshi Hayashi Tadaaki Ooka Mitsuru Shibukawa 《Bioscience, biotechnology, and biochemistry》2013,77(7):1823-1829
Two potent gastroprotective substances against experimental gastric ulcers in rats induced by stress and five of their analogues were isolated from the culture broth of bacterial strain AI-77 which was classified taxonomically as Bacillus pumilus. Physico-chemical properties and pharmacological activities of the seven compounds were examined and compared with each other. 相似文献
3.
Noritsugu Yabe Yutaka Matsuya Isao Yamane Mitsuru Takada 《In vitro cellular & developmental biology. Plant》1986,22(7):363-368
Summary A newly developed, serum-free medium (NYSF-404) selects for antibody-producing hybridomas after fusion of antigen-sensitized
mouse spleen cells with myeloma cell lines P3-X63-Ag8-U1 (P3-U1), P3-X63-Ag8-6.5.3 (Ag8.653), or P3-NSI/1-Ag4-1 (NS-1). Without
the need for hypoxanthine-aminopterinthymidine (HAT) selection of hybrid cells, frequency of hybridoma formation in medium
NYSF-404 is higher (twice) than that in serum- and HAT-containing medium. Colonies developed upon limiting dilution in the
presence of the mortal parent myeloma cells in medium NYSF-404 and pure culture of antibody-secreting cells could be subsequently
established. The results suggest that fusions can be done in serum-free medium and that the clonal growth of hybridomas is
dependent on factors produced by parent myeloma cells under serum-free culture conditions. Such factors seem deficient in
serum- and HAT-containing medium or are masked by serum. 相似文献
4.
Transformation of L cells with virus thymidine kinase genes introduced by red cell-mediated microinjection 总被引:2,自引:0,他引:2
Takashi Iino Mitsuru Furusawa Iwao Furusawa Masuo Obinata 《Experimental cell research》1983,148(2):475-480
Fusion of red cell ghosts containing foreign materials with cells results in the introduction of the materials into the cells (red cell-mediated microinjection). Until now, 'two-step dialysis' has mainly been used for trapping proteins in the ghosts. Large-sized materials such as DNA, however, are rarely trapped in the ghosts, since the holes in the red cell membrane caused by osmotic shock are too small for such materials to pass through. In this study, we improved the trapping technique. Some of the Hind III fragments of lambda phage DNA as well as proteins could be trapped in the ghosts when the mixture of these materials and red cells were frozen at -80 degrees C for a short period followed by quick thawing. Red cell-mediated microinjection using ghosts containing plasmid pBR322 linked with a Herpes simplex viral thymidine kinase (tk) gene brought about transformation of tk-defective L cells, the efficiency of transformation was 1 out of 20 000-60 000 cells fused with the ghosts. 相似文献
5.
Effect of Ethanol on the Sodium and Potassium Conductances of the Squid Axon Membrane 总被引:8,自引:6,他引:2 下载免费PDF全文
The effects of ethanol on squid giant axons were studied by means of the sucrose-gap technique. The membrane action potential height is moderately reduced and the duration sometimes shortened by ethanol in sea water. Voltage clamp experiments showed that ethanol in sea water reduced the maximum membrane conductances for sodium (g'Na) and potassium (g'K). In experiments with multiple application of ethyl alcohol to the same spot of membrane, a reduction of g'Na to 82 per cent and of g'K to 80 per cent of their value in sea water was brought about by 3 per cent ethanol (by volume) while 6 per cent caused a decrease of g'Na to 59 per cent and of g'K to 69 per cent. Ethanol has no significant effect on the steady-state inactivation of gNa (as a function of conditioning membrane potential) or on such kinetic parameters as τh or the time course of turning on gi gNa and gK. It is concluded that ethanol mainly reduces gNa and gK in the Hodgkin-Huxley terminology. 相似文献
6.
Mitsuru Iwata Shoko Iwata Mark A. Everett Bryan B. Fuller 《In vitro cellular & developmental biology. Plant》1990,26(6):554-560
Summary A human foreskin organ culture system has been developed to study the response of human skin to hormonal stimulation. Foreskins
are maintained in culture on floating plastic supports which allows the epidermal surface to be exposed to air while the dermis
is bathed in nutrient medium. Both black and white human foreskins can be maintained in organ culture for at least 1 wk with
no change in the tissue structure or cell viability as determined by histochemical staining and by dopa reaction staining.
Tyrosinase activity in both black and white human foreskin cultures decays markedly during the first 2 d of culture to a new
steady state level which remains stable throughout the culture period. Both black and white foreskin cultures consistently
demonstrate 2- to 10-fold increases in tyrosinase activity when treated with theophylline (1 mM). Approximately 90% of all skin cultures examined showed an increase in enzyme activity when treated with this phosphodiesterase
inhibitor. Dibutyryl cAMP (0.1 mM) and [Nle4, D-phe7]-alpha MSH (10−8
M), were also found to markedly stimulate tyrosinase activity in some skin cultures, whereas alpha-MSH and prostaglandin E1 produced only an inconsistent and small increase in the activity of the enzyme. Histamine (1 μM), vitamin D3 (1 μM), and retinoic acid (1μM) failed to stimulate tyrosinase activity in either white or black foreskin cultures. This hormone-responsive organ culture
system can be utilized to characterize the molecular processes responsible for the regulation of tyrosinase and pigmentation
in human skin.
This work was supported by a research contract from the Oklahoma Center for the Advancement of Science and Technology (OCAST)
and by a research grant from the Presbyterian Health Foundation. 相似文献
7.
John O. Hui John Le Viswanatham Katta Robert Rosenfeld Michael F. Rohde Mitsuru Haniu 《Journal of Protein Chemistry》1996,15(4):351-358
Human neurotrophin-3 (NT-3) is a member of the nerve growth factor (NGF) family of neurotrophic factors, and the recombinant protein is being developed as a therapeutic for neurodegenerative diseases. The final product purity and lot-to-lot variation are monitored routinely by peptide mapping. However, only the N-terminal region of NT-3 was susceptible to proteolysis under native conditions. Complete digestion required that the protein be chemically modified by reduction and S-alkylation prior to proteolysis. Complete proteolytic degradation of the protein was achieved simply by an intial denaturation of NT-3 in 6 M guanidinium chloride (pH 6) for 2 hr at 37°C, followed by a tenfold dilution with the digestion buffer (0.1 M Tris-HCl, 1 mM CaCl2 at pH 7.0) and immediate addition of chymotrypsin at 1% by weight. Direct comparison of the peptide map with an identical aliquot that had been reduced and alkylated also allowed the establishment of the cystine linkages present in NT-3: Cys14 to Cys79, Cys57 to Cys108, and Cys67 to Cys110. This disulfide structure is homologous to the NGF family of neurotrophic factors. 相似文献
8.
Shinichi Kitamura Takashi Hirano Kenichi Takeo Mitsuru Mimura Kanji Kajiwara Bjrn T. Stokke Tsutomu Harada 《International journal of biological macromolecules》1994,16(6)
The conformation and dilute solution properties of (2→1)-β-d-fructan in aqueous solution were studied by gel permeation chromatography, low-angle laser light-scattering photometry, viscometry, small-angle X-ray scattering and electron microscopy. Fractions covering a broad range of weight-average molecular weights (Mw) from 1.49 × 104 to 5.29 × 106 were obtained from a native sample by ultrasonic degradation and fractional precipitation. For Mw < 4 × 104, the intrinsic viscosity [η] varies with Mw0.71, indicating that the fructan chain behaves as a random coil expanded by an excluded-volume effect in this molecular weight region. For Mw > 105, [η] exhibits an unusually weak dependence on Mw and finally becomes almost independent of molecular weight. This behaviour is interpreted in terms of a globular conformation of the high-molecular-weight fructan molecules. Small-angle X-ray-scattering measurements and electron microscopic observations support this interpretation of the values of [η] observed. 相似文献
9.
It has been generally considered that the low productivity of Leguminosae is caused by accumulation in the reproductive organs of a large amount of protein and lipid, since the biochemical costs of synthesizing these compounds is higher than that for carbohydrate. However, we report here on results which show that: the growth efficiencies (dry matter accumulated/ (dry matter accumulated + respiration)) of reproductive organs of Gramineae and Leguminosae were similar; the growth efficiency of rice in the vegetative stage was greater than that of soybean and field bean, regardless of nitrogen application rate; and when 14CO2, 14C-sucrose or 14C-asparagine were introduced to the leaf at the maturation stage, respiratory loss of the introduced 14C was greater in soybean and field bean, especially in the light, than in rice. Thus, it is assumed that the low productivity in Leguminosae is caused by a larger respiratory loss under both dark and light condition in the shoot, and not in the reproductive organs. 相似文献
10.
Kazuo Iwaki Norio Okumura Mitsuru Yamazaki 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1993,619(2)
A rapid and accurate determination of tetracycline antibiotics in human serum by reversed-phase high-performance liquid chromatography with fluorescence detection has been developed, based on protein precipitation in serum. Various reagents for precipitation were investigated, and 24% trichloroacetic acid in methanolic solution gave the maximum recovery (at least 94.3%) and interference-free chromatograms of different three tetracyclines. At a concentration of 0.5 μg/ml, the precision (relative standard deviation) ranged from 1.12 to 1.94%. In the range 0.04–10.0 μg/ml for oxytetracycline and chlorotetracycline and 0.01–10.0 μg/ml for tetracycline, linear responses were observed. The detection limits of this method were 10–35 ng/ml for all three antibiotics. The proposed method was applied to the determination of serum concentrations in subjects receiving tetracycline antibiotics. 相似文献