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121.
利用人粒细胞集落刺激因子(hG-CSF)cDNA3′端非翻译区(3′-UTR)中存在的DraⅠ酶切位点,通过部分酶切与完全酶切,删除3′-UTR不同长度,构建了四种hG-CSFcDNA瞬时重组表达质粒。转染COS-7细胞后,生物活性测定结果提示,hG-CSFcDNA3′-UTR对其表达起负调控作用,其关键性序列位于紧接终止密码子TGA下游的65bp范围内,3′-UTR对hG-CSFcDNA表达的影响与转录水平的差别有一定关系。 相似文献
122.
将编码人单核细胞趋化蛋白-1(MCP-1)的基因亚克隆到大肠杆菌表达载体pEX31A中,在大肠杆菌中表达出MS2/MCP-1融合蛋0白,该表达产物约占菌体总蛋白的15%左右,Westernblot检测表明,表达产物可与MCP-1抗体特异反应。采用琼脂糖平板法进行活性测定表明,表达产物具有明显的单核细胞趋化活性,说明N端融合一段细菌蛋白对MCP-1有无趋化活性可能没有影响。 相似文献
123.
Missense mutations define a restricted segment in the C-terminal domain of phytochrome A critical to its regulatory activity. 总被引:13,自引:4,他引:9
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The phytochrome family of photoreceptors has dual molecular functions: photosensory, involving light signal perception, and regulatory, involving signal transfer to downstream transduction components. To define residues necessary specifically for the regulatory activity of phytochrome A (phyA), we undertook a genetic screen to identify Arabidopsis mutants producing wild-type levels of biologically defective but photochemically active and dimeric phyA molecules. Of eight such mutants identified, six contain missense mutations (including three in the same residue, glycine 727) clustered within a restricted segment in the C-terminal domain of the polypeptide. Quantitative photobiological analysis revealed retention of varying degrees of partial activity among the different alleles--a result consistent with the extent of conservation at the position mutated. Together with additional data, these results indicate that the photoreceptor subdomain identified here is critical to the regulatory activity of both phyA and phyB. 相似文献
124.
Arabidopsis TCH4, regulated by hormones and the environment, encodes a xyloglucan endotransglycosylase. 总被引:17,自引:0,他引:17
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W Xu M M Purugganan D H Polisensky D M Antosiewicz S C Fry J Braam 《The Plant cell》1995,7(10):1555-1567
Adaptation of plants to environmental conditions requires that sensing of external stimuli be linked to mechanisms of morphogenesis. The Arabidopsis TCH (for touch) genes are rapidly upregulated in expression in response to environmental stimuli, but a connection between this molecular response and developmental alterations has not been established. We identified TCH4 as a xyloglucan endotransglycosylase by sequence similarity and enzyme activity. Xyloglucan endotransglycosylases most likely modify cell walls, a fundamental determinant of plant form. We determined that TCH4 expression is regulated by auxin and brassinosteroids, by environmental stimuli, and during development, by a 1-kb region. Expression was restricted to expanding tissues and organs that undergo cell wall modification. Regulation of genes encoding cell wall-modifying enzymes, such as TCH4, may underlie plant morphogenetic responses to the environment. 相似文献
125.
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127.
Previous work showed that in hamster red cells the amiloride-sensitive (AS) Na+ influx of 0.8 mmol/liter cells/hr is not mediated by Na-H exchange as in other red cells, but depends upon intracellular Mg2+ and can be increased by 40-fold by loading cells with Mg2+ to 10 mm. The purpose of this study was to verify the connection of AS Na+ influx with Na-dependent, amiloride-sensitive Mg2+ efflux and to utilize AS Na+ influx to explore that pathway.Determination of unidirectional influx of Na+ and net loss of Mg2+ in parallel sets of cells showed that activation by extracellular [Na+] follows a simple Michaelis-Menten relationship for both processes with a K
m
of 105–107 mm and that activation of both processes is sigmoidally dependent upon cytoplasmic [Mg2+] with a [Mg2+]0.5 of 2.1–2.3 mm and a Hill coefficient of 1.8. Comparison of Vmax for both sets of experiments indicated a stoichiometry of 2 Na: l Mg. Amiloride inhibits Na+ influx and Mg2+ extrusion in parallel (K
i
= 0.3 mm). Like Mg2+ extrusion, amiloride-sensitive Na+ influx shows an absolute requirement for cytoplasmic ATP and is increased by cell swelling. Hence, amiloride-sensitive Na+ influx in hamster red cells appears to be through the Na-Mg exchange pathway.There was no amiloride-sensitive Na+ efflux in hamster red cells loaded with Na+ and incubated with high [Mg2+] in the medium with or without external Na+, nor with ATP depletion. Hence, this is not a simple Na-Mg exchange carrier. 相似文献
128.
S. Chao C. Baysdorfer O. Heredia-Diaz T. Musket G. Xu E. H. Coe Jr 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(6-7):717-721
We report here the results of mapping a set of 92 leaf cDNA clones in maize. The ends of each of these cDNA clones have previously been partially sequenced, and the sequence comparison has revealed the putative function for 28 clones. It is expected that the RFLP map developed using these expressed sequence tags will be of great importance for future maize genome analysis, such as for PCR-based gene mapping or gene function identification.Contribution from the Missouri Agricultural Experiment Station. Journal Series N. 12,019. 相似文献
129.
An influenza A (H1N1) virus, closely related to swine influenza virus, responsible for a fatal case of human influenza. 总被引:8,自引:2,他引:6
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D E Wentworth B L Thompson X Xu H L Regnery A J Cooley M W McGregor N J Cox V S Hinshaw 《Journal of virology》1994,68(4):2051-2058
In July 1991, an influenza A virus, designated A/Maryland/12/91 (A/MD), was isolated from the bronchial secretions of a 27-year-old animal caretaker. He had been admitted to the hospital with bilateral pneumonia and died of acute respiratory distress syndrome 13 days later. Antigenic analyses with postinfection ferret antisera and monoclonal antibodies to recent H1 swine hemagglutinins indicated that the hemagglutinin of this virus was antigenically related to, but distinguishable from, those of other influenza A (H1N1) viruses currently circulating in swine. Oligonucleotide mapping of total viral RNAs revealed differences between A/MD and other contemporary swine viruses. However, partial sequencing of each RNA segment of A/MD demonstrated that all segments were related to those of currently circulating swine viruses. Sequence analysis of the entire hemagglutinin, nucleoprotein, and matrix genes of A/MD revealed a high level of identity with other contemporary swine viruses. Our studies on A/MD emphasize that H1N1 viruses in pigs obviously continue to cross species barriers and infect humans. 相似文献
130.
Rice Triosephosphate Isomerase Gene 5[prime] Sequence Directs [beta]-Glucuronidase Activity in Transgenic Tobacco but Requires an Intron for Expression in Rice 总被引:1,自引:0,他引:1
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In rice (Oryza sativa L.), cytosolic triosephosphate isomerase (TPI) is encoded by a single gene. TPI catalyzes a vital step in glycolysis, and RNA blots showed that the tpi gene is expressed in all vegetative tissues (root, culm, and leaves) and in rice suspension cells. No effect of light on expression was detected, but submergence of rice seedlings resulted in elevated levels of TPI mRNA in roots and culms. The 2767-bp 5[prime] upstream sequence of the tpi gene was fused translationally with the [beta]-glucuronidase (gusA) gene, and the resulting construct, TPI-GUS, was found to express constitutive, high levels of GUS activity in transgenic tobacco (Nicotiana tabacum) plants. However, the same construct yielded no GUS activity in stably transformed rice plants, and RNA blots showed that no GUS mRNA could be detected even though stable integration of functional copies of the construct was confirmed by Southern blot and genomic polymerase chain reaction analyses. Transient assays using particle bombardment yielded high levels of GUS expression from the TPI-GUS construct in tobacco leaves, but essentially no expression in rice, barley, or maize leaves. When the first intron of the tpi gene was included in the construct (TPI-int1-GUS), transient GUS activity was routinely obtained in rice leaves, revealing that the first intron of the rice tpi gene is crucial for its expression in rice. TPI-int1-GUS also directed transient GUS expression in maize and barley leaves, but little or no activity was obtained from this construct in tobacco, tomato, or soybean leaves. These results with the rice tpi promoter are in accordance with mounting evidence that differences in gene expression exist between monocots and dicots. 相似文献