全文获取类型
收费全文 | 15219篇 |
免费 | 806篇 |
国内免费 | 8篇 |
出版年
2022年 | 73篇 |
2021年 | 123篇 |
2020年 | 69篇 |
2019年 | 98篇 |
2018年 | 156篇 |
2017年 | 159篇 |
2016年 | 263篇 |
2015年 | 410篇 |
2014年 | 474篇 |
2013年 | 1253篇 |
2012年 | 837篇 |
2011年 | 895篇 |
2010年 | 516篇 |
2009年 | 487篇 |
2008年 | 838篇 |
2007年 | 921篇 |
2006年 | 903篇 |
2005年 | 950篇 |
2004年 | 1025篇 |
2003年 | 963篇 |
2002年 | 913篇 |
2001年 | 208篇 |
2000年 | 176篇 |
1999年 | 195篇 |
1998年 | 236篇 |
1997年 | 220篇 |
1996年 | 187篇 |
1995年 | 161篇 |
1994年 | 118篇 |
1993年 | 167篇 |
1992年 | 178篇 |
1991年 | 132篇 |
1990年 | 125篇 |
1989年 | 134篇 |
1988年 | 113篇 |
1987年 | 86篇 |
1986年 | 98篇 |
1985年 | 124篇 |
1984年 | 119篇 |
1983年 | 103篇 |
1982年 | 117篇 |
1981年 | 105篇 |
1980年 | 89篇 |
1979年 | 54篇 |
1978年 | 54篇 |
1977年 | 53篇 |
1976年 | 59篇 |
1975年 | 39篇 |
1974年 | 42篇 |
1973年 | 44篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
101.
Yasushi Oda Haruki Nakamura Toshio Yamazaki Kuniaki Nagayama Mayumi Yoshida Shigenori Kanaya Morio Ikehara 《Journal of biomolecular NMR》1992,2(2):137-147
Summary Two-dimensional (2D)1H NMR experiments using deuterium labeling have been carried out to investigate the solution structure of ribonuclease HI (RNase HI) fromEscherichia coli (E. coli), which consists of 155 amino acids. To simplify the1H NMR spectra, two fully deuterated enzymes bearing several prototed amino acids were prepared from an RNase HI overproducing strain ofE. coli grown in an almost fully deuterated medium. One enzyme was selectively labeled by protonated His, He. Val. and Leu. The other was labeled by only protonated His and Ile. The 2D1H NMR spectra of these deuterated R Nase H1 proteins, selectively labeled with protonated amino acids, were much more simple than those of the normally protonated enzyme. The simplified spectra allowed unambiguous assignments of the resonance peaks and connectivities in COSY and NOESY for the side-chain protons. The spin-lattice relaxation times of the side-chain protons of the buried His residue of the deuterated enzyme became remarkably longer than that of the protonated enzyme. In contrast, the relaxation times of the side-chain protons of exposed His residues remained essentially unchanged. 相似文献
102.
103.
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
104.
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
105.
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
106.
NMR studies for identification of dI:dG mismatch base-pairing structure in DNA. 总被引:4,自引:4,他引:0
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
One- and two-dimensional NMR experiments have been undertaken to investigate deoxyinosine:deoxyguanosine (dI:dG) base pairing in a self-complementary dodecadeoxyribonucleotide, d(C1-G2-C3-I4-A5-A6-T7-T8-G9-G10-G11-G12) (designated IG-12), duplex. The NMR data indicate formation of a dI(syn):dG(anti) base pair in a B-DNA helix. This unusual base pairing results in altered NOE patterns between the base protons (H8 and H2) of the I4 residue and the sugar protons of its own and the 5'-flanking C3 residues. The dI(syn):dG(anti) base pair is accommodated in the B-DNA duplex with only a subtle distortion of the local conformation. Identification of the dI:dG base pairing in this study confirms that a hypoxanthine base can form hydrogen-bonded base pairs with all of the four normal bases, C, A, T, and G, in DNA. 相似文献
107.
108.
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
109.
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
110.
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