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1.
Identification of an Rts1 DNA fragment conferring temperature-dependent instability to vector plasmids 总被引:4,自引:0,他引:4
The multiphenotypic drug resistance factor Rts1 expresses a temperature-dependent instability characteristic. This plasmid was digested with the restriction enzyme BamHI. A DNA fragment with a molecular weight of 5.6 MDa (the H fragment) was inserted into plasmid pBR322 (pFK896) or into pSC105 (pYH156) at the BamHI site. These plasmids were unstable at 42 degrees C but stable at 32 degrees C. A restriction-enzyme map of the H fragment was constructed and the instability phenotype (Tdi) was localized to a DNA fragment with 0.5 MDa molecular weight. The temperature-dependent loss of the unstable plasmid pFK896 is abrupt and no gradual plasmid loss of this multicopy recombinant plasmid is observed. The possibility that the Tdi phenotype is due to overgrowth of R- cells was eliminated. 相似文献
2.
R Parrilla K Okawa K O Lindros U J Zimmerman K Kobayashi J R Williamson 《The Journal of biological chemistry》1974,249(15):4926-4933
3.
Rho-associated kinase, a novel serine/threonine kinase, as a putative target for small GTP binding protein Rho. 总被引:23,自引:2,他引:21
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T Matsui M Amano T Yamamoto K Chihara M Nakafuku M Ito T Nakano K Okawa A Iwamatsu K Kaibuchi 《The EMBO journal》1996,15(9):2208-2216
The small GTP binding protein Rho is implicated in cytoskeletal responses to extracellular signals such as lysophosphatidic acid to form stress fibers and focal contacts. Here we have purified a Rho-interacting protein with a molecular mass of approximately 164 kDa (p164) from bovine brain. This protein bound to GTPgammaS (a non-hydrolyzable GTP analog).RhoA but not to GDP.RhoA or GTPgammaS.RhoA with a mutation in the effector domain (RhoAA37).p164 had a kinase activity which was specifically stimulated by GTPgammaS.RhoA. We obtained the cDNA encoding p164 on the basis of its partial amino acid sequences and named it Rho-associated kinase (Rho-kinase). Rho-kinase has a catalytic domain in the N-terminal portion, a coiled coil domain in the middle portion and a zinc finger-like motif in the C-terminal portion. The catalytic domain shares 72% sequence homology with that of myotonic dystrophy kinase and the coiled coil domain contains a Rho-interacting interface. When COS7 cells were cotransfected with Rho-kinase and activated RhoA, some Rho-kinase was recruited to membranes. Thus it is likely that Rho-kinase is a putative target serine/threonine kinase for Rho and serves as a mediator of the Rho-dependent signaling pathway. 相似文献
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1. Phospholipase D [EC 3.1.4.4] from Streptomyces hachijoensis was purified about 570-fold by column chromatography on DEAE-cellulose and Sephadex G-50 followed by isoelectric focusing. 2. The purified preparation was found to be homogeneous both by immunodiffusion and polyacrylamide disc gel electrophoresis. 3. The isoelectric point was found to be around pH 8.6 and the molecular weight was about 16,000. 4. The enzyme has maximal activity at pH 7.5 at 37 degrees. The optimal temperature is around 50 degrees at pH 7.5, using 20 min incubation. 5. The enzyme was stable at 50 degrees for 90 min. At neutral pH, between 6 and 8, the enzyme retained more than 95% of its activity on 24 hr incubation at 25 degrees. However, the enzyme lost 80% of its activity under the same conditions at pH 4.0. 6. The enzyme was stimulated slightly by Ca2+, Mn2+, and Co2+, and significantly by Triton X-100 and ethyl ether. It was inhibited by Sn2+, Fe2+, Fe3+, Al3+, EDTA, sodium dodecyl sulfate, sodium cholate, and cetylpyridinium chloride. 7. This phospholipase D hydrolyzes phosphatidylethanolamine, phosphatidylcholine, cardiolipin, sphingomyelin, phosphatidylserine, and lysophosphatidylcholine, liberating the corresponding bases. 8. The Km value was 4mM, determined with phosphatidylethanolamine as a substrate. 相似文献
7.
Articular lesions were studied in the extremities and vertebral column in murine erysipelas infection. Pathologic alterations found in those different joint systems were essentially the same in character. In the early stage of infection, phlegmon-like inflammation was observed in the periarticular loose connective tissue. Abundant fine Gram-positive bacilli were free in inflamed edematous tissue. Then, active serofibrinous exudation and emigration of leukocytes occurred in the synovial membrane, articular cartilage, annulus fibrosus, and nucleus pulposus with the development of the disease. These exudative changes disappeared gradually. Instead of them, proliferation of synovial cells accompanied by dense infiltration with lymphocytes and plasma cells became conspicuous. At the same time, chondro- and osteoclastic activity occurred in the joints of the extremities or regions adjacent to the intervertebral disk. Capsular and subchondral pannus often resulted from an excessive proliferation of granulation tissue. In the late stage, they synovial membrane and granulation tissue underwent fibrosis, adhesion and ossification. Finally, these changes resulted in fibrous or bony ankylosis and deformation. Gram-positive bacilli were hardly detected in histological preparations derived from animals in the chronic stage when proliferative change was predominant in the articular lesions. 相似文献
8.
Misa Ohno Yuto Togashi Kyoko Tsuda Kazuaki Okawa Minori Kamaya Masayoshi Sakaguchi Yasusato Sugahara Fumitaka Oyama 《PloS one》2013,8(6)
Chitinase hydrolyzes chitin, which is an N-acetyl-D-glucosamine polymer that is present in a wide range of organisms, including insects, parasites and fungi. Although mammals do not contain any endogenous chitin, humans and mice express two active chitinases, chitotriosidase (Chit1) and acidic mammalian chitinase (AMCase). Because the level of expression of these chitinases is increased in many inflammatory conditions, including Gaucher disease and mouse models of asthma, both chitinases may play important roles in the pathophysiologies of these and other diseases. We recently established a quantitative PCR system using a single standard DNA and showed that AMCase mRNA is synthesized at extraordinarily high levels in mouse stomach tissues. In this study, we applied this methodology to the quantification of chitinase mRNAs in human tissues and found that both chitinase mRNAs were widely expressed in normal human tissues. Chit1 mRNA was highly expressed in the human lung, whereas AMCase mRNA was not overexpressed in normal human stomach tissues. The levels of these mRNAs in human tissues were significantly lower than the levels of housekeeping genes. Because the AMCase expression levels were quite different between the human and mouse stomach tissues, we developed a quantitative PCR system to compare the mRNA levels between human and mouse tissues using a human-mouse hybrid standard DNA. Our analysis showed that Chit1 mRNA is expressed at similar levels in normal human and mouse lung. In contrast, the AMCase expression level in human stomach was significantly lower than that expression level observed in mouse stomach. These mRNA differences between human and mouse stomach tissues were reflecting differences in the chitinolytic activities and levels of protein expression. Thus, the expression level of the AMCase in the stomach is species-specific. 相似文献
9.
Tanida Kotomi Shimada Mihoko Khor Seik-Soon Toyoda Hiromi Kato Kayoko Kotorii Nozomu Kotorii Tatayu Ariyoshi Yu Kato Takao Hiejima Hiroshi Ozone Motohiro Uchimura Naohisa Ikegami Azusa Kume Kazuhiko Kanbayashi Takashi Imanishi Aya Kamei Yuichi Hida Akiko Wada Yamato Kuroda Kenji Miyamoto Masayuki Hirata Koichi Takami Masanori Yamada Naoto Okawa Masako Omata Naoto Kondo Hideaki Kodama Tohru Inoue Yuichi Mishima Kazuo Honda Makoto Tokunaga Katsushi Miyagawa Taku 《Sleep and biological rhythms》2022,20(1):137-148
Sleep and Biological Rhythms - Idiopathic hypersomnia (IH) is a rare sleep disorder characterized by excessive daytime sleepiness, great difficulty upon awakening, and prolonged sleep time. In... 相似文献
10.
T Okawa H Ichimal T Ishida M Kawata S Kaguba K Mamba T Makita 《Cell biology international reports》1989,13(6):547-553
To estimate the possibility that biological degradable starch microspheres (DMS) activate abdominal or intraperitoneal macrophages (IMP), two sizes of DMS (Spherex, Pharmacia, Sweden) were injected into the peritoneum of the ICR mice of 4 to 8 weeks of age. Three days after the injection, peritoneal fluid was collected and incubated for one hour at 37 degrees C under 5% CO2. The cells which adhered to the petri dish were IMP, to which DMS was added for 18 hrs. The cultured IMP were observed by scanning electron microscope (SEM) and the ratio of the active type to the total number of IMP was counted as an index of the effect of DMS to IMP. The activation effect of DMS on the incubated IMP was significant in the group which was cultured with 2 microns DMS after the 45 microns DMS injection. That indicated the possible DMS function as a potential IMP activating factor (MAF). 相似文献