全文获取类型
收费全文 | 187633篇 |
免费 | 14791篇 |
国内免费 | 13655篇 |
出版年
2024年 | 358篇 |
2023年 | 2285篇 |
2022年 | 5059篇 |
2021年 | 8704篇 |
2020年 | 5880篇 |
2019年 | 7322篇 |
2018年 | 7156篇 |
2017年 | 5350篇 |
2016年 | 7567篇 |
2015年 | 11014篇 |
2014年 | 13015篇 |
2013年 | 13892篇 |
2012年 | 16764篇 |
2011年 | 15219篇 |
2010年 | 9511篇 |
2009年 | 8407篇 |
2008年 | 9893篇 |
2007年 | 8917篇 |
2006年 | 7884篇 |
2005年 | 6242篇 |
2004年 | 5420篇 |
2003年 | 4851篇 |
2002年 | 4211篇 |
2001年 | 3648篇 |
2000年 | 3387篇 |
1999年 | 3305篇 |
1998年 | 1888篇 |
1997年 | 1996篇 |
1996年 | 1809篇 |
1995年 | 1651篇 |
1994年 | 1597篇 |
1993年 | 1207篇 |
1992年 | 1731篇 |
1991年 | 1381篇 |
1990年 | 1042篇 |
1989年 | 962篇 |
1988年 | 810篇 |
1987年 | 715篇 |
1986年 | 617篇 |
1985年 | 640篇 |
1984年 | 359篇 |
1983年 | 335篇 |
1982年 | 231篇 |
1981年 | 192篇 |
1980年 | 162篇 |
1979年 | 189篇 |
1978年 | 127篇 |
1977年 | 118篇 |
1975年 | 112篇 |
1974年 | 126篇 |
排序方式: 共有10000条查询结果,搜索用时 78 毫秒
941.
武汉东湖磷含量的变动及其分布 总被引:2,自引:0,他引:2
本文总结了武汉东湖水体中磷含量(均以PO4计算)的周年逐月季节和年际变动及其分布上的差异(1973-1985年)。按面积加权法计算总磷的平均含量为0.244毫克/升(1983-1985年),总溶解磷和溶解活性磷的平均含量分别为0.121毫克/升和0.051毫克/升(1981-1984年),总磷和总溶解磷周年中出现两次高峰含量,即春季(3-5月)和夏末秋初(8-9月)。低含量出现在水温最低的冬季(12-2月),周年中溶解活性磷高峰含量出现在冬末春初(1-3月),低含量多数出现在春天夏初(5-7月)。东湖水体中磷含量平面分布有明显的差异,而垂直分布表层和底层差异小,各种形态磷的组成中颗粒磷所占比较最大(1983-1984年平均值),平均占总磷63.4%,溶解非活性磷所占比较最小,平均占总磷12.0%。 相似文献
942.
943.
Cloning of genes governing the deoxysugar portion of the erythromycin biosynthesis pathway in Saccharopolyspora erythraea (Streptomyces erythreus). 总被引:29,自引:21,他引:8
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
J Vara M Lewandowska-Skarbek Y G Wang S Donadio C R Hutchinson 《Journal of bacteriology》1989,171(11):5872-5881
Genes that govern the formation of deoxysugars or their attachment to erythronolide B and 3 alpha-mycarosyl erythronolide B, intermediates of the biosynthesis of the 14-membered macrolide antibiotic erythromycin, were cloned from Saccharopolyspora erythraea (formerly Streptomyces erythreus). Segments of DNA that complement the eryB25, eryB26, eryB46, eryC1-60, and eryD24 mutations blocking the formation of erythronolide B or 3 alpha-mycarosyl erythronolide B, when cloned in Escherichia coli-Streptomyces shuttle cosmids or plasmid vectors that can transform S. erythraea, were located in a ca. 18-kilobase-pair region upstream of the erythromycin resistance (ermE) gene. The eryC1 gene lies just to the 5' side of ermE, and one (or possibly two) eryB gene is approximately 12 kilobase pairs farther upstream. Another eryB gene may be in the same region, while an additional eryB mutation appears to be located elsewhere. The eryD gene lies between the eryB and eryC1 genes and may regulate their function on the basis of the phenotype of an EryD- mutant. 相似文献
944.
A part of the GTP gamma S-binding activity in murine thymocyte membranes was found to have affinity to a concanavalin A (Con A)-Sepharose column. The material was identified as Gi (inhibitory GTP-binding protein) on the basis of the molecular weight and by islet activating protein-dependent ADP-ribosylation and anti-alpha i (alpha subunit of Gi) immunoblotting. However, when the membranes prepared from Con A-stimulated thymocytes were used, no GTP gamma S-binding activity was detected in the Con A-bound fraction, suggesting that Gi physically and specifically associated with Con A acceptors dissociates upon Con A stimulation. Furthermore, another GTP gamma S-binding protein (25 kDa), which is quite similar to a novel phosphoinositide-specific phospholipase C (PI-PLC)-associated G protein in calf thymocytes (Wang, P., Toyoshima, S., & Osawa, T. (1988) J. Biochem. 103, 137-142), was detected among the Con A-Sepharose-bound proteins with the chemical cross-linking technique. When the 40 kDa and 25 kDa G proteins associated with Con A receptor(s) were isolated and their direct effects on the activity of partially purified PI-PLC as to phosphatidylinositol 4,5-bisphosphate hydrolysis were examined, the 25 kDa G protein was found to enhance the PI-PLC activity more effectively. On the other hand, pretreatment of cells with islet-activating protein completely abolished the inhibitory effect of Con A on the prostaglandin E1 and isoproterenol-induced increases of cellular cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
945.
Phosphorylation of highly purified growth hormone receptors by a growth hormone receptor-associated tyrosine kinase 总被引:5,自引:0,他引:5
C Carter-Su J R Stubbart X Y Wang S E Stred L S Argetsinger J A Shafer 《The Journal of biological chemistry》1989,264(31):18654-18661
We have shown previously that growth hormone (GH) promotes the phosphorylation of its receptor on tyrosyl residues (Foster, C. M., Shafer, J. A., Rozsa, F. W., Wang, X., Lewis, S. D., Renken, D. A., Natale, J. E., Schwartz, J., and Carter-Su, C. (1988) Biochemistry 27, 326-334). In the present study, we investigated the possibility that a tyrosine kinase is specifically associated with the GH receptor. GH-receptor complexes were first partially purified from GH-treated 3T3-F442A fibroblasts, a GH-responsive cell, by immunoprecipitation using anti-GH antiserum. 35S-Labeled proteins of Mr = 105,000-125,000 were observed in the immunoprecipitate from GH-treated cells labeled metabolically with 35S-amino-acids. These proteins were not observed in immunoprecipitates from cells not exposed to GH or when non-immune serum replaced the anti-GH antiserum, consistent with the proteins being GH receptors. GH receptors appeared to be phosphorylated, as evidenced by the presence of 32P-labeled bands, comigrating with the 105-125 kDa 35S-labeled proteins, in the immunoprecipitate of GH-treated cells labeled metabolically with [32P]Pi. When partially purified GH receptor preparation was incubated with [gamma-32P]ATP (7-15 microM) for 10 min at 30 degrees C in the presence of MnCl2, a protein of Mr = 121,000 was phosphorylated exclusively on tyrosyl residues. As expected for the GH receptor, this protein was not observed in immunoprecipitates when cells had not been treated with GH nor when non-immune serum replaced the anti-GH antiserum. GH-receptor complexes were also purified to near homogeneity by sequential immunoprecipitation with phosphotyrosyl-binding antibody followed by anti-GH antiserum. When cells were labeled metabolically with 35S-amino acids, the 35S label migrated almost exclusively as an Mr = 105,000-125,000 protein. This protein also incorporated 32P into tyrosyl residues when incubated in solution with [gamma-32P]ATP. These results show that highly purified GH receptor preparations undergo tyrosyl phosphorylation, suggesting that either the GH receptor itself is a tyrosine kinase or is tightly associated with a tyrosine kinase. 相似文献
946.
We have isolated for the first time two kinds of endo-beta-N-acetylglucosaminidases (E-beta-GNases) simultaneously from human kidney. E-beta-GNase 1 was purified by water extraction, ammonium sulfate fractionation, and chromatography on Sephadex-G-200, DEAE-Sephadex, concanavalin A-Sepharose and Hypatite C columns. After the DEAE-Sephadex step, 107 units of E-beta-GNase 1 with a specific activity of 0.53 units/mg was obtained and after hydroxyapatite column, the enzyme recovery was 26 units with a specific activity of 10.4 units/mg. This enzyme hydrolyzed the high mannose-type asparaginylglycopeptide efficiently and had little activity toward the complex-type glycopeptide. This enzyme had an pH optimum at about 4.5 and was not inhibited by acetate ion. The Asn residue in a glycopeptide appeared not to be an important recognition site for E-beta-GNase 1 to express its activity because the acetylation or the dansylation of Asn residues as well as the elimination of Asn residue from the glycopeptide did not change the susceptibility of the oligosaccharide to E-beta-GNase 1. E-beta-GNase 2 was purified by water extraction, ammonium sulfate fractionation, and chromatography on Sephadex G-200, DEAE-Sephadex, concanavalin A-Sepharose, and Mono S columns. This enzyme was purified about 110-fold with 6.6% recovery. E-beta-GNase 2 was found to be a novel type of E-beta-GNase that hydrolyzed both the high mannose-type and the complex-type oligosaccharide with chitobiosyl group at the reducing end and without the Asn. E-beta-GNase 2 activity was found to be dependent on a L-aspartamido-beta-D-N-acetylglucosamine amidohydrolase (Asn-GNase) for the hydrolysis of asparaginylglycopeptide. Asn-GNase cleaved off the Asn residue from the glycopeptide, and the resulting oligosaccharide was hydrolyzed by E-beta-GNase 2. Because the acetylation or the dansylation of Asn residue in a glycopeptide rendered the glycopeptide resistant to Asn-GNase, the use of the modified asparaginylglycopeptide could not reveal the existence of E-beta-GNase 2 activity. The pH optimum of E-beta-GNase was found to be about 3.5. Like beta-hexosaminidases, this enzyme was inhibited by acetate ion, suggesting the recognition of GlcNAc moiety by this enzyme. 相似文献
947.
948.
C W Slife E Wang R Hunter S Wang C Burgess D C Liotta A H Merrill 《The Journal of biological chemistry》1989,264(18):10371-10377
Long-chain (sphingoid) bases may serve as another category of "lipid second messenger" because they inhibit protein kinase C and affect multiple cellular functions. Free sphingosine has been found in rat liver (Merrill, A. H., Jr., Wang, E., Mullins, R. E., Jamison, W. C. L., Nimkar, S., and Liotta, D. C. (1988) Anal. Biochem. 171, 373-381); hence, this study determined if liver plasma membranes contain free long-chain bases and have the ability to form them from endogenous enzymes and substrates. Isolated plasma membranes contained 0.45 nmol of sphingosine/mg of protein which, based on the recovery of the membranes, was equivalent to 3.5 +/- 1.2 nmol/g of liver and at least half of the total free sphingosine in liver. When the membranes were incubated at 37 degrees C, the amount increased at an initial rate of 5-25 pmol/min/mg, resulting in a 2-3-fold increase over an hour. Sphingosine formation required divalent cations, was optimal at neutral to alkaline pH, and was temperature-dependent. Activities with these characteristics were not identified in microsomes or lysosomes (lysosomal activities with acidic pH optima were detected, however); hence, they appear to reflect a separate plasma membrane system. Sphingosine formation was stimulated by ceramides either added exogenously or formed endogenously by treating the membranes with sphingomyelinase (but not endoglycoceramidase). Sphingomyelin hydrolysis to ceramide was also observed during incubation of the plasma membranes alone. Some of the properties of this system resembled the neutral sphingomyelinase and ceramidase activities of liver. While the physiological significance of this endogenous sphingosine is not known, this system has the appropriate subcellular location to provide sphingosine as a participant in signal transduction. 相似文献
949.
950.
A human muscle adenine nucleotide translocator gene has four exons, is located on chromosome 4, and is differentially expressed 总被引:8,自引:0,他引:8
K Li C K Warner J A Hodge S Minoshima J Kudoh R Fukuyama M Maekawa Y Shimizu N Shimizu D C Wallace 《The Journal of biological chemistry》1989,264(24):13998-14004