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161.
A human protein selected for interference with Ras function interacts directly with Ras and competes with Raf1. 总被引:13,自引:8,他引:5 下载免费PDF全文
The overexpression of some human proteins can cause interference with the Ras signal transduction pathway in the yeast Saccharomyces cerevisiae. The functional block is located at the level of the effector itself, since these proteins do not suppress activating mutations further downstream in the same pathway. We now demonstrate, with in vivo and in vitro experiments, that the protein encoded by one human cDNA (clone 99) can interact directly with yeast Ras2p and with human H-Ras protein, and we have named this gene rin1 (Ras interaction/interference). The interaction between Ras and Rin1 is enhanced when Ras is bound to GTP. Rin1 is not able to interact with either an effector mutant or a dominant negative mutant of H-Ras. Thus, Rin1 displays a human H-Ras interaction profile that is the same as that seen for Raf1 and yeast adenylyl cyclase, two known effectors of Ras. Moreover, Raf1 directly competes with Rin1 for binding to H-Ras in vitro. Unlike Raf1, however, the Rin1 protein resides primarily at the plasma membrane, where H-Ras is localized. These data are consistent with Rin1 functioning in mammalian cells as an effector or regulator of H-Ras. 相似文献
162.
Deformation and flow of red blood cells in a synthetic lattice: evidence for an active cytoskeleton. 总被引:3,自引:0,他引:3 下载免费PDF全文
We introduce the use of microfabrication techniques to construct on a silicon wafer a synthetic capillary bed with 2.5- to 4-micron (mu)-wide channels. Establishment of a fluid pressure gradient allowed us to observe simultaneously using optical microscopy hundreds of cells flowing through the bed at physiological speeds. We find a large distribution of mobilities among red cells flowing through the structure; smaller channels provide a greater impedance to flow than larger ones, indicating that kinetic drag variations provide the origin of the distribution. The mobility of a particular cell is not correlated with the cell diameter but appears to be inversely correlated with intracellular calcium concentration of the cell, as determined by fluorescence of the calcium-binding dye fluo-3 AM. Also, we are able to use the parallel processing nature of our arrays to observe isolated events where the rigidity of the red cell seems to change suddenly over several orders of magnitude as it blocks a channel in the array. 相似文献
163.
原生质体电融合酵母多倍体生理特性的研究 总被引:5,自引:0,他引:5
对原生质体电融合技术获得的11株酵母多倍体融合株,进行了一系列生理特性的分析比较.结果发现,在电融合过程中,融合株的细胞体积及DNA含量并不随着核倍性呈线性递增.而是有其特殊的变化规律。当糖化酵母sta1、sta2和sta3在细胞核中各自纯合时,明显表现出表达的剂量效应。其中sta1、sta2纯合的融合株.在YEPS培养中,GA分泌可被淀粉大量诱导,表现出一定的二次生长特性。从分析结果推测,在亲株5301-14D及HU-TY—1A中可能有对GA分泌不利的因素。 相似文献
164.
兔感觉神经特异蛋白的纯化及稳定性观察 总被引:2,自引:0,他引:2
以兔脊神经节及背根纤维为材料,通过制备匀浆,DEAE-Sephacel阴离子交换层析,高压液相凝胶过滤层析分离纯化了感觉神经特异蛋白29 ku,并进行了该蛋白的稳定性观察. 相似文献
165.
抗白粉病小麦染色体组型的分子标记与生化标记分析 总被引:1,自引:0,他引:1
应用与小麦第六同源群有关的分子和生化标记,包括DNA探针pSc5·3H3和pSR167以及同工酶Est-5和a-Amy-1,对来自六倍体小黑麦Beagle与普通小麦科冬58杂交后代F1花粉植株的抗白粉病株系M24.M09及M17进行了分析。结果表明,M24、M09及M17不同程度地含有黑麦染色体成分,而且电泳谱带差别较大,据此推断,M09为6RL的易位系。因此,生化和分子标记不仅可以用于确定外源片段的存在,而且可以帮助确定染色体组型和外源片段的位置 相似文献
166.
Identification of new mutations in the Cu/Zn superoxide dismutase gene of patients with familial amyotrophic lateral sclerosis. 总被引:6,自引:0,他引:6 下载免费PDF全文
A Pramatarova D A Figlewicz A Krizus F Y Han I Ceballos-Picot A Nicole M Dib V Meininger R H Brown G A Rouleau 《American journal of human genetics》1995,56(3):592-596
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder affecting motor neurons. Although most cases of ALS are sporadic, approximately 10% are inherited as an autosomal dominant trait. Mutations in the Cu/Zn superoxide dismutase gene (SOD 1) are responsible for a fraction of familial ALS (FALS). Screening our FALS kindreds by SSCP, we have identified mutations in 15 families, of which 9 have not been previously reported. Two of the new mutations alter amino acids that have never been implicated in FALS. One of them affects a highly conserved amino acid involved in dimer contact, and the other one affects the active-site loop of the enzyme. These two mutations reduce significantly SOD 1 enzyme activity in lymphoblasts. Our results suggest that SOD 1 mutations are responsible for > or = 13% of FALS cases. 相似文献
167.
Construction and use of a nontoxigenic strain of Pseudomonas aeruginosa for the production of recombinant exotoxin A. 下载免费PDF全文
To express recombinant forms of Pseudomonas aeruginosa exotoxin A in high yield, we have developed a nontoxigenic strain of P. aeruginosa derived from the hypertoxigenic strain PA103. The nontoxigenic strain, designated PA103A, was produced by the excision marker rescue technique to replace the toxA structural gene in PA103 with an insertionally inactivated toxA gene. The PA103A strain (ToxA-) was used subsequently as the host strain for the expression and production of several recombinant versions of exotoxin A, and the results were compared with exotoxin A production in other P. aeruginosa and Escherichia coli strains. Use of the PA103A strain transformed with the high-copy-number pRO1614 plasmid bearing various toxA alleles resulted in final purification yields of exotoxin A averaging 23 mg/liter of culture. By comparison, exotoxin A production in other expression systems and host strains yields approximately 1/4 to 1/10 as much toxin. 相似文献
168.
Le Van Phung Yuki Han Shiro Oka Hisako Hotta Michael D. Smith Prapit Theeparakun Eiko Yabuuchi Ikuya Yano 《FEMS immunology and medical microbiology》1995,12(3-4):259-264
Abstract The serodiagnosis of melioidosis is commonly performed with tests using protein or polysaccharide as antigen. However, due to the low sensitivity, specificity and difficulty in the preparation of the antigens, more simple, precise and reproducible diagnostic tests were required. A purified glycolipid antigen (GL) which is a specific lipid component of Burkholderia pseudomallei has been used in an ELISA. With this antigen, specific immunoglobulin G (IgG) was detected in 49 out of 50 melioidosis sera. IgG was also detected in 2 out of 185 (Japanese) and 16 out of 181 (Vietnamese) control sera. Thus, the sensitivity was 98.0%, and specificity was 98.9% and 91.1% in the Japanese and Vietnamese sera, respectively. When the ELISA and indirect haemagglutination (IHA) tests were combined, a sensitivity of 100% and specificity of 97.8% were achieved. The advantages of the glycolipid antigen are ease of preparation, stability, high sensitivity and specificity. 相似文献
169.
Preparation and structural characterization of large heparin-derived oligosaccharides 总被引:4,自引:2,他引:2
Pervin Azra; Gallo Cindy; Jandik Kenneth A; Han Xue-Jun; Linhardt Robert J. 《Glycobiology》1995,5(1):83-95
Porcine mucosal heparin was partially depolymerized with heparinlyase I and then fractionated into low-molecularweight (<5000)and high-molecular-weight (>5000) oligosaccharides by pressurefiltration. The high-molecular-weight oligosaccharide mixture({small tilde}50 wt% of the starting heparin) also containedintact heparin. This intact polymer complicates oligosacsharidepurification. Thus, the low-molecular-weight fraction was usedto prepare homogeneous oligosaccharides for structural characterization.The low-molecular-weight oligosaccharide mixture was first fractionatedby low pressure gel permeation chromatography into size-uniformmixtures of disaccharides, tetrasaccharides, hexasaccharides,octasaccharides, decasaccharides, dodecasaccharides, tetradecasaccharidesand higher oligosaccharides. Each size-fractionated mixturewas then purified on the basis of charge by repetitive semi-preparativestrong-anion-exchange high-performance liquid chromatography.This approach has led to the isolation of 14 homogeneous oligosaccharidesfrom disaccharide to tetradecasaccharide. The purity of theseheparin-derived oligosaccharides was determined by gradientpolyacrylamide gel electrophoresis, analytical strong-anion-exchangehigh-performance liquid chromatography, capillary electrophoresisand one-dimensional nuclear resonance spectroscopy. The structureof these oligosaccharides was established using 600 MHz two-dimensionalnuclear resonance spectroscopy . The spectral methods used includedhomonuclear correlation spectroscopy, nuclear Overhauser effectspectroscopy and heteronuclear multiple quantum coherence spech-clscopy.The 1H/1H connectivities of the protons of each sugar residuein an oligosaccharide were established by two-dimensional homonuclearcorrelation spectroscopy, while 1H/13C assignments were madeusing 1H inverse detection. One- and two-dimensional nuclearresonance spectroscopic analysis of these heparin oligosaccharidesshowed two closely related groups of heparin-oligosaccharidesare afforded by enzymatic depolymerization of heparin. One groupis fully sulphated, having the structures 相似文献
170.
四川地区幼儿和学龄前儿童的鼻部测量 总被引:2,自引:0,他引:2
本文报告1116例四川地区幼儿和学龄前儿童(2-7岁)鼻部9项指标的测量均数,性差及年龄发育特点。性差:仅鼻凹鼻底距4-6.5岁等少数指标部分年龄段男女性间出现显著性划异(男>女)。此外各项指标的绝大多数年龄段男女性间无显著性差异。年龄发育:9项测量指标中7项的生长曲线随年产长而上升,数值随年龄增大,并有1-2个发育高峰;提示鼻部发育具有阶段性;2项指标的曲线随年龄增长变化较小。4项指标男女性的曲 相似文献