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101.
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
102.
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
103.
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
104.
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
105.
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
106.
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109.
A microorganism that produces glucose isomerase was isolated from soil and identified as a strain of Streptomyces flavogriseus. The organism produced a large quantity of glucose isomerase when grown on straw hemicellulose, xylan, xylose, and H2SO4 hydrolysate of ryegrass straw. The organism produced glucose isomerase both intra- and extra-cellularly. The highest level of intracellular glucose isomerase (3.5 U/ml) was obtained in about 36 h by a culture grown on straw hemicellulose; the extracellular enzyme (1.5 U/ml) appeared in cultures grown for about 72 h. About equal levels of enzyme were produced in cultures grown on straw hemicellulose, xylan, xylose, and H2SO4 hydrolysate of straw, but production of the enzyme was drastically reduced when the organism was grown on other carbon sources. As a nitrogen source, corn steep liquor produced the best results. Soy flour extract, yeast extract, and various peptones also were adequate substrates for glucose isomerase production. Addition of Mg2+, Mn2+, or Fe2+ to the growth medium significantly enhanced enzyme production. The organism, however, did not require Co2+, which is commonly required by microorganisms used in the production of glucose isomerase. 相似文献
110.
The cellular immune response (MIF, E-rosette formation and changes in nucleolar morphology of lymphocytes) was followed as
related to age and antigenic stimulation. MIF in healthy infants increased from the 2nd to the 12th week of life as compared
with the first week, probably due to BCG vaccination. The total and active E-rosette formation did not change during the whole
period of investigation. Ring-shaped nucleoli increased gradually from the second week of life. Active nucleoli increased
up to the 4th week,i.e. after BCG vaccination and then slowly decreased. Micronucleoli being high in the first week, decreased during 24 weeks of
life. After artificial colonization of the intestine the production of MIF was slightly lower in colonized infants than in
controls from the 2nd to the 12th week. The other parameters followed were not influenced by colonization. 相似文献