全文获取类型
收费全文 | 108082篇 |
免费 | 8417篇 |
国内免费 | 8910篇 |
出版年
2024年 | 240篇 |
2023年 | 1417篇 |
2022年 | 3256篇 |
2021年 | 5511篇 |
2020年 | 3777篇 |
2019年 | 4679篇 |
2018年 | 4428篇 |
2017年 | 3238篇 |
2016年 | 4591篇 |
2015年 | 6676篇 |
2014年 | 7844篇 |
2013年 | 8296篇 |
2012年 | 9978篇 |
2011年 | 8974篇 |
2010年 | 5545篇 |
2009年 | 4970篇 |
2008年 | 5710篇 |
2007年 | 5130篇 |
2006年 | 4454篇 |
2005年 | 3491篇 |
2004年 | 2969篇 |
2003年 | 2719篇 |
2002年 | 2272篇 |
2001年 | 1866篇 |
2000年 | 1694篇 |
1999年 | 1670篇 |
1998年 | 1035篇 |
1997年 | 1001篇 |
1996年 | 941篇 |
1995年 | 821篇 |
1994年 | 787篇 |
1993年 | 617篇 |
1992年 | 818篇 |
1991年 | 617篇 |
1990年 | 467篇 |
1989年 | 443篇 |
1988年 | 354篇 |
1987年 | 344篇 |
1986年 | 266篇 |
1985年 | 286篇 |
1984年 | 156篇 |
1983年 | 161篇 |
1982年 | 99篇 |
1981年 | 85篇 |
1980年 | 60篇 |
1979年 | 77篇 |
1977年 | 59篇 |
1975年 | 56篇 |
1974年 | 52篇 |
1973年 | 56篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
981.
Naoki Sugawara Dan Li Chieko Sugawara Hirotsugu Miyake 《Biological trace element research》1995,49(2-3):161-169
Fischer rats were a fed diet supplied with copper chloride (150–600 ppm) for 60 d from weaning. Serum (glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) activities were increased with the increase of Cu concentration in the diet. Biliary excretion of Cu was related to the dietary Cu level. Depositions of hepatic and renal Cu were also related to the dietary Cu level in a dose-dependent manner. In particular, hepatic (155.2±13.3 μg/g) and renal (44.9±4.4 μg/g) Cu concentrations increased abruptly in the Cu-600 ppm group. In the liver, about 60% of Cu was distributed in the soluble fraction (100,000 g supernatant). In the Cu-600 ppm group, 25% of cystosolic Cu was bound to metallothionein (MT). Our results suggest that chronic exposure to Cu appears to have a deleterious effect on the hepatic function, and further, that even in rats with normal biliary Cu excretion, clearance of Cu from the liver may be marginal when dietary Cu is near the 600-ppm level. Although Cu is an essential nutrient, an overload of Cu should be avoided. 相似文献
982.
Human cytomegalovirus neutralizing antibody-resistant phenotype is associated with reduced expression of glycoprotein H. 总被引:8,自引:4,他引:4
下载免费PDF全文
![点击此处可从《Journal of virology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
We have characterized a neutralizing antibody-resistant mutant human cytomegalovirus (HCMV) obtained from a patient treated with a human monoclonal antiglycoprotein H (gH; unique long region 75) antibody. This virus exhibited resistance to several different neutralizing anti-gH murine monoclonal antibodies (MAbs), as well as to a polyvalent anti-gH serum. The resistant phenotype was unstable and could be maintained only by passage of plaque-purified virus under neutralizing MAb selection. In the absence of a MAb, the resistant phenotype reverted to a neutralizing antibody-sensitive phenotype within one passage. The predicted amino acid sequences of gH from the MAb-resistant and -susceptible parent viruses were identical. Biochemical analysis of the MAb-resistant and -susceptible parent viruses revealed a marked decrease of gH expression in the envelope of the MAb-resistant virus. Furthermore, propagation of the virus in various MAb concentrations resulted in the production of extracellular virions with various levels of resistance to the neutralizing activity of the MAb. These results suggest a mechanism for the generation of neutralizing antibody-resistant viruses which could evade host-derived antiviral antibody responses. In addition, our findings indicate that the stoichiometry of gH in the envelope of infectious HCMV virions is not rigidly fixed and therefore offer a simple explanation for production of phenotypic variants of HCMV through an assembly process in which the content of gH in the envelope of progeny virions varies randomly. 相似文献
983.
Essential role of the Vp2 and Vp3 DNA-binding domain in simian virus 40 morphogenesis. 总被引:4,自引:3,他引:1
下载免费PDF全文
![点击此处可从《Journal of virology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Both a DNA-binding domain and a Vp1 interactive determinant have been mapped to the carboxy-terminal 40 residues of the simian virus 40 (SV40) minor capsid proteins, Vp2 and Vp3 (Vp2/3), with the last 13 residues being necessary for these activities. The role of this DNA-binding domain in SV40 morphogenesis and the ability to separate these two signals were investigated by mutagenesis and assessment of the activity and viability of the mutants. The carboxy-terminal 40 residues of Vp2/3 were expressed as a polyhistidine fusion protein, and five basic residues at the extreme carboxy terminus (Vp3 residues K226, R227, R228, R230, and R233) were mutagenized. The wild-type fusion protein bound DNA with a Kd of 3 x 10(-8) identical to that of the full-length Vp3. Mutant proteins containing either one to three or four amino acid substitutions bound DNA 4- to 7-fold or 20- to 30-fold less well, respectively, than the wild-type protein did. The most severe point mutants showed residual DNA binding similar to that of a truncated protein which lacks the entire 13 carboxy-terminal residues. All of the point mutants were able to interact with Vp1, indicating that the two signals within this region are mediated by different residues. When the mutations were placed into the context of the viral DNA and introduced into cells, all the structural proteins were expressed and localized correctly. Not all, however, were viable: mutant genomes whose Vp2/3 bound DNA with intermediate affinities formed plaques just as well as wild-type SV40 DNA did, but three mutants showing greatly reduced DNA binding failed to form plaques at all. These results are consistent with the hypothesis that Vp2/3 plays an essential role in SV40 virion assembly in the nucleus. 相似文献
984.
Michael E. Pacold Fred J. Stevens Dong Li Louise E. Anderson 《Photosynthesis research》1995,43(2):125-130
Light activation of NADP-linked glyceraldehyde-3-P dehydrogenase involves reductive cleavage of a disulfide bond. We have proposed that the inactivating disulfide locks the two domains of the enzyme, preventing catalysis, and we have tentatively identified the two critical cysteine residues in the chloroplast enzyme (D. Li, F.J. Stevens, M. Schiffer and L.E. Anderson (1994) Biophys J. 67: 29–35). We reasoned that if activation of this enzyme involves these cysteines that enzymes lacking one or both should be active in the dark and insensitive to reductants. One of these cysteines is present in the enzymes from Anabaena variabilis and Synechocystis PCC 6803 but the other is not. Consistent with the proposed mechanism, glyceraldehyde-3-P dehydrogenase is not affected by DTT-treatment in extracts of either of these cyanobacteria. Fructosebisphosphatase is DTT-activated in extracts of both of these cyanobacteria and glucose-6-P dehydrogenase is inactivated in Synechocystis, as in higher plant chloroplasts. Apparently reductive modulation is possible in these cyanobacteria but glyceraldehyde-3-P dehydrogenase is not light activated. 相似文献
985.
986.
We have purified and characterized poly(A) polymerases (PAPs) from Pisum sativum, Brassica juncea, and Zea mays. Through chromatography on DEAE-Sepharose and heparin-Sepharose, these PAPs copurified as a single enzyme along with RNPs that could provide RNA substrates for the enzyme. More extensive purification by chromatography on MonoQ resulted in the resolution of the PAPs into as many as three fractions. One of these (PAP-I) contained a 43-kDa polypeptide immunologically related to the yeast PAP, and two others (PAP-II and PAP-III) contained RNAs that could serve as substrates for polyadenylation. These fractions by themselves possessed little PAP activity, but mixtures containing combinations of these displayed substantial activity. Similar PAP factors (PAP-I and PAP-III) were identified after fractionation of extracts prepared from Brassica juncea and Zea mays. The factors from one plant were completely interchangeable with those from different plants. We conclude that the poly(A) polymerases present in vegetative plant tissues consist of more than one component. In this respect, they are substantially different from other reported plant, mammalian, and yeast PAPs. 相似文献
987.
988.
Nutrients and heavy metal contamination of plants and sediments in Futian mangrove forest 总被引:2,自引:1,他引:1
An ecological survey was carried out to determine the levels of nutrients and heavy metals in the sediments and leaf tissues of two dominant mangrove plant species, Kandelia candel and Aegiceras corniculatum, in Futian mangrove forest, Shenzhen, the People's Republic of China. The spatial and seasonal variations of these elements were also investigated. The results show that there was no major difference between two sampling sites 150 m apart. In both sites, the sediment concentrations of total and NH4
+-N, total and extractable P, total and extractable K, total organic carbon were consistently higher in the landward locations and decreased gradually towards the sea. The sediment sample collected at the seaward edge of the mangrove plant community had the lowest levels of nutrient and organic matter. The vertical variations (from the land to the sea) of sediment heavy metals were less obvious and no particular trend could be identified. Extremely high contents of Cu, Cd, Pb, Cr and Zn were found at certain locations, suggesting the occurrence of some local contamination. The mean total metal concentrations in sediments decreased in the order Mn > Zn > Cu > Cr = Pb > Cd for the sample locations. Most of the heavy metals were not in a bioavailable form as the concentrations of extractable metals were relatively low (< 1% of total metals). Pb, Cr and Cd were not detected in leaf samples. Leaf C, N, P and K contents were similar between the two species and no significant difference was found among locations, although A. corniculatum seemed to have lower Mn concentrations than K. candel. With reference to temporal variations, no significant difference in sediment concentrations of some nutrients and metals was found between the spring and autumn seasons. 相似文献
989.
A 22-kb segment of chromosomal DNA from Enterococcus faecalis OG1RF containing the pyrimidine biosynthesis genes pyrC and pyrD was previously detected as complementing Escherichia coli pyrC and pyrD mutations. In the present study, it was found that the E. faecalis pyrimidine biosynthetic genes in this clone (designated pKV48) are part of a larger cluster resembling that seen in Bacillus spp. Transposon insertions were isolated at a number of sites throughout the cluster and resulted in loss of the ability to complement E. coli auxotrophs. The DNA sequences of the entire pyrD gene of E. faecalis and selected parts of the rest of the cluster were determined, and computer analyses found these to be similar to genes from Bacillus subtilis and Bacillus caldolyticus pyrimidine biosynthesis operons. Five of the transposon insertions were introduced back into the E. faecalis chromosome, and all except insertions in pyrD resulted in pyrimidine auxotrophy. The prototrophy of pyrD knockouts was observed for two different insertions and suggests that E. faecalis is similar to Lactococcus lactis, which has been shown to possess two pyrD genes. A similar analysis was performed with the purL gene from E. faecalis, contained in another cosmid clone, and purine auxotrophs were isolated. In addition, a pool of random transposon insertions in pKV48, isolated in E. coli, was introduced into the E. faecalis chromosome en masse, and an auxotroph was obtained. These results demonstrate a new methodology for constructing defined knockout mutations in E. faecalis. 相似文献
990.
The twisting and writhing during growth of single-cell filaments of Bacillus subtilis which lead to macrofiber formation was studied in both left- and right-handed forms of strains FJ7 and RHX. Filament bending, touching, and loop formation (folding), followed by winding up into a double-strand fiber, were documented. Subsequent folds that produced multistrandedness were also examined. The rate of loop rotation during winding up was measured for 26 loops from 16 clones. In most cases, the first loop formed turned at a lower rate than those produced by the following cycles of folding. The sequence of folding topologies differed in FJ7 and RHX strains and in left- versus right-handed structures. Right-handed FJ7 routinely gave rise to four-stranded helices at the second fold, whereas left-handed FJ7 and both left-handed and right-handed forms of RHX made structures with predominantly two double-stranded helical regions. Left-handed RHX structures frequently produced second folds within the initial loop itself, resulting in T- or Y-shaped fibers. Sixteen cases in which the initial touch of a filament to itself produced a loop that snapped open before it could wind up into a double-strand fiber were found. The snap motions were used to obtain estimates of the forces generated by helical growth of single filaments and to investigate theoretical models involving the material properties of cell filaments. In general, the mechanical behavior of growing single-cell filaments and fibers consisting of two-, three-, or four-strand helices was similar to that described for larger, mature, multifilament macrofibers. The behavior of multicellular macrofibers can be understood, therefore, in terms of individual cell growth. 相似文献