全文获取类型
收费全文 | 14632篇 |
免费 | 740篇 |
国内免费 | 8篇 |
出版年
2022年 | 62篇 |
2021年 | 126篇 |
2020年 | 62篇 |
2019年 | 88篇 |
2018年 | 146篇 |
2017年 | 152篇 |
2016年 | 259篇 |
2015年 | 395篇 |
2014年 | 452篇 |
2013年 | 1197篇 |
2012年 | 798篇 |
2011年 | 860篇 |
2010年 | 502篇 |
2009年 | 477篇 |
2008年 | 816篇 |
2007年 | 902篇 |
2006年 | 888篇 |
2005年 | 940篇 |
2004年 | 1016篇 |
2003年 | 944篇 |
2002年 | 891篇 |
2001年 | 182篇 |
2000年 | 149篇 |
1999年 | 181篇 |
1998年 | 229篇 |
1997年 | 216篇 |
1996年 | 182篇 |
1995年 | 153篇 |
1994年 | 125篇 |
1993年 | 164篇 |
1992年 | 156篇 |
1991年 | 118篇 |
1990年 | 111篇 |
1989年 | 115篇 |
1988年 | 99篇 |
1987年 | 76篇 |
1986年 | 81篇 |
1985年 | 94篇 |
1984年 | 105篇 |
1983年 | 95篇 |
1982年 | 109篇 |
1981年 | 106篇 |
1980年 | 88篇 |
1979年 | 45篇 |
1978年 | 52篇 |
1977年 | 46篇 |
1976年 | 51篇 |
1975年 | 31篇 |
1974年 | 43篇 |
1973年 | 42篇 |
排序方式: 共有10000条查询结果,搜索用时 453 毫秒
91.
92.
The permeability of bacterial outer membranes was assayed by coupling the influx of highly hydrophobic probes, 3-oxosteroids, with their subsequent oxidation catalysed by 3-oxosteroid delta 1-dehydrogenase, expressed from a gene cloned from Pseudomonas testosteroni. In Salmonella typhimurium producing wild-type lipopolysaccharide, the permeability coefficients for uncharged steroids were 0.45 to 1 x 10(-5) cm s-1, and the diffusion appeared to occur mainly through the lipid bilayer domains of the outer membrane. These rates are one or two magnitudes lower than that expected for their diffusion through the usual biological membranes. The permeation rates were markedly increased (up to 100 times) when the lipopolysaccharide leaflet was perturbed either by adding deacylpolymyxin or by introducing mutations leading to the production of deep rough lipopolysaccharides. An amphiphilic, negatively charged probe, testosterone hemisuccinate, penetrated much more slowly than the uncharged steroids. Study of various Gram-negative species revealed that P. testosteroni, Pseudomonas acidovorans, and Acinetobacter calcoaceticus showed higher outer membrane permeability to steroid probes and higher susceptibility to hydrophobic agents such as fusidic acid, novobiocin and crystal violet relative to S. typhimurium and Escherichia coli. 相似文献
93.
Takashi Saito Kensuke Futatsugi Daisuke Miki Hiroshi Suzuki Kiyoshi Yasukawa 《Biotechnology Techniques》1992,6(4):365-370
Summary Murine interleukin-6 (mIL-6) was expressed inEscherichia coli as human growth hormone (hGH) fusion protein. The products were cleaved by thrombin to liberate mIL-6. Monoclonal and polyclonal
antibodies specific to mIL-6 were prepared by immunizing rats with mIL-6 thus obtained. ELISA for the quantitation of mIL-6
was also established, which could detect mIL-6 in a quantity as low as 2 ng/ml. 相似文献
94.
Summary A cell-associated pullalanase (-dextrin 6-glucanohydrolase, EC 3.2.1.41) of an extreme thermophile, Bacillus flavocaldarius KP 1228, was purified to homogeneity. The molecular weight and isoelectric point were estimated to be about 55 000 and 7.0, respectively. The N-terminal sequence was Ala-Try-Tyr-Glu-Gly-Ala-Phe-Phe-Tyr-Gln-Ile-Phe-Pro-Asp-Tyr-Phe-Phe-Tyr-Ala-Gly-. The enzyme was most active at pH 6.3. The activities for 5% pullulan and 5% soluble starch were maximal at 75–80° C and at 80–85° C, respectively. The enzyme was stable up to 90° C for 10 min at pH 6.8. The enzyme had no antigenic determinants shared with pullulanases from the mesophiles Klebsiella pneumoniae and B. acidopullulyticus NCIB 11647. A comparison of amino acid composition demonstrated that the proline content increased greatly in a linear fashion with the rise in thermostability in the order K. pneumoniae B. acidopullulyticus B. flavocaldarius enzymes, as found with Bacillus oligo-1,6-glucosidases.Presented in part at the Annual Meeting of the Agricultural Chemical Society of Japan at Tokyo, April 2, 1987 (Abstracts, p 91)Offprint requests to: Y. Suzuki 相似文献
95.
Xenopus M phase MAP kinase: isolation of its cDNA and activation by MPF. 总被引:53,自引:15,他引:38
下载免费PDF全文
![点击此处可从《The EMBO journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Y Gotoh K Moriyama S Matsuda E Okumura T Kishimoto H Kawasaki K Suzuki I Yahara H Sakai E Nishida 《The EMBO journal》1991,10(9):2661-2668
MAP kinase is activated and phosphorylated during M phase of the Xenopus oocyte cell cycle, and induces the interphase-M phase transition of microtubule dynamics in vitro. We have carried out molecular cloning of Xenopus M phase MAP kinase and report its entire amino acid sequence. There is no marked change in the MAP kinase mRNA level during the cell cycle. Moreover, studies with an anti-MAP kinase antiserum indicate that MAP kinase activity may be regulated posttranslationally, most likely by phosphorylation. We show that MAP kinase can be activated by microinjection of MPF into immature oocytes or by adding MPF to cell-free extracts of interphase eggs. These results suggest that MAP kinase functions as an intermediate between MPF and the interphase-M phase transition of microtubule organization. 相似文献
96.
97.
Teruo Okasora Mitsumasa Nagase Hiroshi Kawachi Katsuyuki Matsui Michiaki Orikasa Tetsuo Morioka Ichiroh Yamazaki Takashi Oite Fujio Shimizu 《Virchows Archiv. B, Cell pathology including molecular pathology》1991,60(1):41-46
Change in the localization of the antigen recognized by the proteinuria-inducing monoclonal antibody (MA) 5-1-6 in experimental nephrosis was studied by indirect and biotin-avidin immunofluorescence, and immunoperoxidase at light and electron microscopical levels. The proteinuric state was induced by the administration of the aminonucleoside of puromycin (PAN) or adriamycin. The antigen decreased in quantity and/or its distribution changed with an increase in the amount of protein excreted in both experimental models. Recovery from the alterations observed during the development and proteinuria appeared to occur when PAN-induced proteinuria subsided. This antigenic molecule may thus be essential for maintaining the normal permselectivity of glomerular capillary walls. 相似文献
98.
Association of Lps gene with natural resistance of mouse macrophages against Legionella pneumophila 总被引:3,自引:0,他引:3
Shin-ichi Yoshida Yoshitaka Goto Hiroshi Miyamoto Hironobu Fujio Yasuo Mizuguchi 《FEMS microbiology letters》1991,89(1):51-56
We have cloned a 1.6-kb region of chromosomal DNA from Thermoplasma acidophilum into Escherichia coli using as a probe part of the Methanococcus vannielii fus-gene. The sequence of the clone was highly homologous to part of the corresponding Methanococcus vannielii gene. By chromosome walking, a 4.7-kb EcoRI fragment containing the complete gene was isolated. Nucleotide sequencing revealed an open reading frame of 2196 nucleotides. The deduced amino acid sequence contains the known peptide sequence around the ADP-ribosylation site of T. acidophilum elongation factor 2, which unequivocally confirms that the fus-gene has been cloned. The amino acid sequence was compared to that of hamster and E. coli, as well as to known archaebacterial EF-2 sequences. 相似文献
99.
Incorporation of fatty acids by Streptococcus mutans 总被引:1,自引:0,他引:1
Masaru Sato Hironori Tsuchiya Hideki Tani Kohji Yamamoto Ryozo Yamaguchi Hiroshi Nitta Nobutake Kanematsu Isamu Namikawa Nobuhiko Takagi 《FEMS microbiology letters》1991,81(1):117-121
In a series of investigations into the cariogenicity of Streptococcus mutans, we studied the incorporation of exogenous fatty acids with reference to glucosyltransferase secretion and membrane fatty acid changes. When cells were grown with different fatty acids, both saturated and unsaturated fatty acids were readily incorporated into the membrane lipids and were biotransformed and elongated preferentially to the longer 16- and 18-carbon-chain fatty acids. This incorporation and chain-elongation led to significant changes in fatty acids composition. By adding fatty acids to the medium, it was possible to appropriately modify the degree of unsaturation and the relative ratio between specific fatty acids in the membrane lipids of S. mutans. 相似文献
100.
A group of circular dichroism (CD) active phospholipids has been synthesised, in which one or both acyl chains has been replaced with a cinnamoyl or azobenzene chromophore-containing acid. Studies on the structure, CD activity and thermodynamic property of liposome membranes composed of CD active phospholipids were carried out. CD active liposomes were found to be stable, normal liposomes of approximately 550 A diameter based on the electron micrograph and dynamic light scattering, and to have thermodynamic property similar to the conventional phospholipid membranes without serious perturbation by aromatic bulk groups based on DSC. Liposomes composed of phospholipid having two trans-azobenzene chromophores showed an extremely large CD enhancement even well above Tc. This CD enhancement was drastically changed by the presence of cis-azobenzene chromophore and cis-cis isomer content after irradiation was higher than the theoretical value, suggesting the importance of interchromophore interaction in the liposome membranes. 相似文献