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21.
Expression in Escherichia coli of chemically synthesized gene for the human immune interferon. 总被引:6,自引:0,他引:6
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S Tanaka T Oshima K Ohsuye T Ono A Mizono A Ueno H Nakazato M Tsujimoto N Higashi T Noguchi 《Nucleic acids research》1983,11(6):1707-1723
A 454 base pair fragment of double stranded DNA consisting of a gene for a human immune interferon (hIFN-gamma), initiation and termination signals plus appropriate restriction endonuclease sites, was totally synthesized. The synthesis involved preparation of 62 oligodeoxyribonucleotides by rapid, solid phase procedures, and enzymatic ligation of the oligonucleotides. This synthetic gene was expressed in E. coli under the control of the lac UV5 promoter. The product has antiviral activity which was acid labile and completely neutralized by antiserum to hIFN-gamma but not by antiserum to hIFN-alpha or hIFN-beta. Molecular weight of hIFN-gamma produced by E. coli was estimated to be about 32,000 and 17,000 by gel filtration and SDS-polyacrylamide gel electrophoresis respectively. 相似文献
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23.
Purification and Chemical Composition of Reticulate Bodies of the Meningopneumonitis Organisms 总被引:23,自引:13,他引:10
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Reticulate bodies of the meningopneumonitis (MP) microorganism were purified from L cells 18 hr after infection by the combination of differential centrifugation in 30% sucrose solution and potassium tartrate density gradient centrifugation. It was ascertained by electron microscopy that purified preparations of reticulate bodies obtained were almost entirely free of host-cell components and of infectious elementary bodies of MP microorganisms. Purified reticulate bodies were easily disrupted by mechanical agitation, and it was observed in shadowed preparation that ribosome-like particles 15 mmu in diameter were scattered from broken reticulate bodies. In shadowed preparations, reticulate bodies were found to range in size from 1.0 to 1.6 mu in diameter, but in cross-section the range was 0.5 to 1.0 mu. In these preparations, the purified reticulate bodies were irregular in shape, round or oval, and were composed of rather homogenous, amorphous, or reticulate material with moderate density. Some particles exhibited a less-dense internal structure, in which a coarse fibrous reticulum was seen. Chemical fractionation of (32)P-labeled purified reticulate bodies showed that they contained three times more ribonucleic acid (RNA) than deoxyribonucleic acid, with the RNA being composed primarily of 21S, 16S, and 4S RNA. No infectivity of purified reticulate bodies could be demonstrated. 相似文献
24.
25.
Comparative study of the sugar chains of gamma-glutamyltranspeptidases purified from human hepatocellular carcinoma and from human liver 总被引:2,自引:0,他引:2
K Yamashita K Totani Y Iwaki I Takamisawa N Tateishi T Higashi Y Sakamoto A Kobata 《Journal of biochemistry》1989,105(5):728-735
The sugar chains of gamma-glutamyltranspeptidases, purified from human hepatoma and from normal human liver, were released quantitatively as oligosaccharides by hydrazinolysis. Comparative study of their structures revealed that the following structural alterations are induced by hepatocyte carcinogenesis: 1) high mannose type sugar chains are detected in the hepatoma enzyme but not in the normal liver enzyme; 2) abnormal biantennary sugar chains containing C-2,4 outer chain branches newly appeared; 3) the total amounts of tri- and tetraantennary sugar chains containing C-2,6 outer chain branches increased up to three times. 相似文献
26.
S W Chensue I G Otterness G I Higashi C S Forsch S L Kunkel 《Journal of immunology (Baltimore, Md. : 1950)》1989,142(4):1281-1286
The present study examined the potential role of IL-1 and TNF in granuloma formation. Mice were given i.v. injections of Schistosoma mansoni eggs or Sephadex beads to induce synchronous immune T cell-mediated (hypersensitivity type) or nonimmune (foreign-body type) granulomas, respectively. Granuloma macrophages isolated at 2, 4, 8, 16, and 32 days of granuloma formation were evaluated for their capacity to produce IL-1 and TNF in response to 1 microgram/ml LPS. This was related to circulating levels of the acute phase protein, serum amyloid P (SAP) and expression of Ia Ag by monocytes and macrophages. Macrophages from nonimmune bead lesions were generally weak producers of IL-1 and TNF. In contrast, those from T cell-mediated egg lesions produced significant levels of both monokines. Moreover, there was a clear pattern of sequential monokine production such that IL-1 was produced in greatest amounts early (2 to 4 days), whereas TNF was produced later (8 to 16 days). Levels of SAP showed an initial sharp rise following particle embolization, then decreased rapidly in bead injected animals. However, mice with immune granulomas showed a prolonged elevation in SAP levels that corresponded to the period of maximal IL-1 production (2 to 4 days). Macrophage/monocyte Ia Ag expression was greatest at 8 to 16 days, corresponding to the period of TNF production. Bead injected animals showed low levels of Ia expression over the study period. These findings suggest that IL-1 may be important in the early recruitment stages of granuloma formation while TNF may take part in later maintenance or effector functions. The extent of production of both is likely influenced by the local or systemic milieu of lymphokines. 相似文献
27.
Interactive effects of salinity, nitrogen and sulphur on the organic solutes in Spartina alterniflora leaf blades 总被引:4,自引:0,他引:4
Colmer Timothy D.; Teresa W-M. Fan; Luchli Andr; Higashi Richard M. 《Journal of experimental botany》1996,47(3):369-375
Glycinebetaine, proline, asparagine, sucrose, glucose, and dimethylsulphoniopropionate(DMSP) were the major organic solutes in Spartina alternifloraleaf blades. To investigate the physiological role(s) of thesesolutes, the effects of salinity, nitrogen, and sulphur treatmentson leaf blade solute levels were examined. Glycinebetaine wasthe major organic solute accumulated in leaf blades grown at500 mol m3 NaCl, although asparagine and proline alsoaccumulated when the supply of nitrogen was sufficient. Thesesolutes may play a role in osmotic adjustment. In contrast,DMSP levels either did not change or were reduced in responseto the 500 mol m3 NaCl treatment. Furthermore, elevatednitrogen supply decreased leaf blade DMSP levels, which wasopposite to the response of glycinebetaine, proline, and asparagine.A 1000-fold increase in external sulphate concentration hadno effect on the leaf blade levels of DMSP, glycinebetaine,proline, or asparagine. These findings suggest that the majorphysiological role of DMSP in S. alterniflora leaf blades isnot for osmotic adjustment, even under conditions of nitrogendeficit and excess sulphur. Instead, DMSP which was presentat 45130 µmol g1 dry weight, may play arole as a constitutive organic osmoticum. Key words: Spartina alterniflora, dimethylsulphoniopropionate, glycinebetaine, nitrogen, salinity 相似文献
28.
Ryousuke Takahashi Katsumi Kawamura Jianguo Hu †Michiyuki Hayashi Takeo Deguchi 《Journal of neurochemistry》1996,67(2):525-529
Abstract: To study the level of ciliary neurotrophic factor (CNTF) in human nervous tissues, we developed a sensitive enzyme-linked immunoassay using a specific antibody against human CNTF. This method allowed us to detect as little as 0.3 ng/ml of human CNTF with good linearity and accuracy. Using this method, CNTF levels were determined in human sciatic nerves obtained at autopsy from 21 amyotrophic lateral sclerosis (ALS) patients and 48 subjects who had died of other neurological diseases. CNTF genotypes were also determined. The results indicated that CNTF levels were high in the normal homozygotes and approximately halved in the heterozygote subjects. There was, however, no significant difference in CNTF levels in the sciatic nerves between ALS and other neurological disease patients, indicating that the CNTF level was mainly determined by its genotypes and that the level in the sciatic nerves was not reduced in ALS patients. 相似文献
29.
Neutral red (NR) in the culture medium entered the vacuolesof a green alga, Micrasterias pinnatifida, at a higher rateat pH 8 than at pH 5. NR remained soluble in vacuoles of cellscultured at pH 5, while it precipitated and formed granulesin cells cultured at pH 8. The vacuoles of cells cultured atpH 8 contained fibrils, but those of cells cultured at pH 5did not. The amount of NR that entered the cells was markedlyreduced by the addition to the medium of nigericin at 10-5M,monensin at 10-5M, bafilo-mycin A1 at 10-5M, or ammonium chlorideat 50 mM. The formation of NR granules in vacuoles were stronglyinhibited and the disorganization of NR granules were acceleratedby the addition of nigericin at 10-5M, or bafilomycin A1 at10-5M to the culture medium. The possibility is discussed thatNR which enters vacuoles might become positively charged (NRH+)by protons brought into vacuoles by proton pumps and that NRH+might combine with some negatively charged macromolecules toform aggregates or granules. (Received April 18, 1996; Accepted May 27, 1996) 相似文献
30.
Sub Sunoo Katsumi Asano Fumiyuki Mitsumori 《European journal of applied physiology and occupational physiology》1996,74(4):305-310
We measured ATP, phosphocreatine (PCr), inorganic phosphate (Pi), and the intracellular pH in rat hindlimb muscles during submaximal isometric exercise with various O2 deliveries using31P nuclear magnetic resonance spectroscopy (31P NMR) to evaluate changes in energy metabolism in relation to O2 availability. Delivery of O2 to muscles was altered by controlling the fractional concentration of inspired oxygen (F
IO2) at 0.50, 0.28, 0.21, 0.11 and 0.08 with monitoring partial pressure of oxygen and carbon dioxide, and bicarbonate at the femoral artery. The steady-state ratio of PCr : (PCr + Pi) during exercise decreased as a function ofF
IO2 even at 0.21. Significant acidification of the intracellular pH during exercise occurred at 0.08F
IO2. Change in the PCr : (PCr + Pi) ratio demonstrated that the oxidative capacity, i.e. the maximal rate of the oxidative phosphorylation reaction, in muscle was not limited by O2 delivery at 0.50F
IO2, but was significantly limited at 0.21F
IO2 or below. Change in the intracellular pH at 0.08F
IO2 could be interpreted as an increase in lactate, suggesting activation of glycolysis. Correlation between the PCr : (PCr + Pi) ratio and the intracellular pH revealed the existence of a critical PCr : (PCr + Pi) ratio and pH for glycolysis activation at around 0.4 and 6.7, respectively. 相似文献