全文获取类型
收费全文 | 108016篇 |
免费 | 8384篇 |
国内免费 | 9010篇 |
专业分类
125410篇 |
出版年
2024年 | 240篇 |
2023年 | 1417篇 |
2022年 | 3260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 | 2968篇 |
2003年 | 2719篇 |
2002年 | 2272篇 |
2001年 | 1866篇 |
2000年 | 1694篇 |
1999年 | 1669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 | 466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
171.
A major autolysin of Pseudomonas aeruginosa: subcellular distribution, potential role in cell growth and division and secretion in surface membrane vesicles. 总被引:3,自引:1,他引:2 下载免费PDF全文
A 26-kDa murein hydrolase is the major autolysin of Pseudomonas aeruginosa PAO1, and its expression can be correlated with the growth and division of cells in both batch and synchronously growing cultures. In batch cultures, it is detected primarily during the mid-exponential growth phase, and in synchronous cultures, it is detected primarily during the cell elongation and division phases. Immunogold labeling of thin sections of P. aeruginosa using antibodies raised against the 26-kDa autolysin revealed that it is associated mainly with the cell envelope and in particular within the periplasm. It is also tightly bound to the peptidoglycan layer, since murein sacculi, isolated by boiling 4% sodium dodecyl sulfate treatment, could also be immunogold labeled. Since division is due to cell constriction in this P. aeruginosa strain (septa are rarely seen), we cannot comment on the autolysin's contribution to septation, although constriction sites were always heavily labeled. Some labeling was also found in the cytoplasm, and this was thought to be due to the de novo synthesis of the enzyme before translocation to the periplasm. Interestingly, the autolysin was also found to be associated with natural membrane vesicles which blebbed from the surface during cell growth; the enzyme is therefore part of the complex makeup of these membrane packages of secreted materials (J. L. Kadurugamuwa and T. J. Beveridge, J. Bacteriol. 177:3998-4008, 1995). The expression of these membrane vesicles was correlated with the expression of B-band lipopolysaccharide. 相似文献
172.
Cell proteins bind specifically to West Nile virus minus-strand 3' stem-loop RNA. 总被引:3,自引:2,他引:1 下载免费PDF全文
The first 96 nucleotides of the 5'noncoding region (NCR) of West Nile virus (WNV) genomic RNA were previously reported to form thermodynamically predicted stem-loop (SL) structures that are conserved among flaviviruses. The complementary minus-strand 3' NCR RNA, which is thought to function as a promoter for the synthesis of plus-strand RNA, forms a corresponding predicted SL structure. RNase probing of the WNV 3' minus-strand stem-loop RNA [WNV (-)3' SL RNA] confirmed the existence of a terminal secondary structure. RNA-protein binding studies were performed with BHK S100 cytoplasmic extracts and in vitro-synthesized WNV (-)3' SL RNA as the probe. Three RNA-protein complexes (complexes 1,2, and 3) were detected by a gel mobility shift assay, and the specificity of the RNA-protein interactions was confirmed by gel mobility shift and UV-induced cross-linking competition assays. Four BHK cell proteins with molecular masses of 108, 60, 50, and 42 kDa were detected by UV-induced cross-linking to the WNV (-)3' SL RNA. A preliminary mapping study indicated that all four proteins bound to the first 75 nucleotides of the WNV 3' minus-strand RNA, the region that contains the terminal SL. A flavivirus resistance phenotype was previously shown to be inherited in mice as a single, autosomal dominant allele. The efficiencies of infection of resistant cells and susceptible cells are similar, but resistant cells (C3H/RV) produce less genomic RNA than congenic, susceptible cells (C3H/He). Three RNA-protein complexes and four UV-induced cross-linked cell proteins with mobilities identical to those detected in BHK cell extracts with the WNV (-)3' SL RNA were found in both C3H/RV and C3H/He cell extracts. However, the half-life of the C3H/RV complex 1 was three times longer than that of the C3H/He complex 1. It is possible that the increased binding activity of one of the resistant cell proteins for the flavivirus minus-strand RNA could result in a reduced synthesis of plus-strand RNA as observed with the flavivirus resistance phenotype. 相似文献
173.
Purification and in vitro phosphorylation of Myxococcus xanthus AsgA protein. 总被引:2,自引:2,他引:0 下载免费PDF全文
The deduced amino acid sequence of the Myxococcus xanthus AsgA protein contains an N-terminal domain that is homologous to the receiver of response regulators and a C-terminal domain that is homologous to the transmitter of histidine protein kinases. We overexpressed affinity-tagged AsgA in Escherichia coli, purified the recombinant protein, and showed that AsgA has autokinase activity in vitro. The results of chemical-stability assays suggest that AsgA is phosphorylated on a histidine and provide no evidence for transfer of the phosphoryl group to the conserved aspartate of the receiver domain. 相似文献
174.
Adherence Patterns and DNA Probe Types of Escherichia coli Isolated from Diarrheal Patients in China
Jian-Guo Xu Bo-Qun Cheng Yan-Ping Wu Li-Bao Huang Xin-He Lai Bing-Yang Liu Xing-Zu Lo Hun-Fen Li 《Microbiology and immunology》1996,40(2):89-97
One hundred and seventy-two strains of Escherichia coli isolated from diarrheal patients in Beijing, P. R. China, were analyzed for plasmid DNA profile, HEp-2 cell adherence ability and reactivity to 10 previously described DNA probes. They had not been recognized as pathogenic E. coli in China. Of the 110 strains tested, 76 (69%) contained one or multiple large plasmids. Of the 71 strains with the large plasmids 64 could adhere to HEp-2 cells. Of the 172 strains, 102 (59.3%) were hybridized with at least one of the 10 probes. Of those, seven strains hybridized with enteroaggregative E. coli (EAggEC) probe. Their serotypes were O128 (two strains), O6 (one strain), and O111 (one strain). Three strains were untypable. Six and three strains were hybridized with enteropathogenic E. coli (EPEC) attaching and effacing genes (eae) or EPEC adherence factor (EAF) probe, respectively. Two non-O157: H7 strains hybridized with enterohemorrhagic E. coli (EHEC) probe. Seventy-two strains (41.9%) hybridized with shiga-like toxin 2 or 1 (SLT2 or SLT1) probes. Among the SLT1 or SLT2 probe-positive strains, 54 hybridized with invasive (INV) plasmid probe developed for identification of enteroinvasive E. coli (EIEC) and Shigella species. The INV and SLT probe-positive strains might represent a new variety of verotoxin-producing E. coli (VTEC). 相似文献
175.
1983年我国报道了从γ-射线处理的“矮杆齐”大麦中得到了一株黄绿色的突变体1832C[1]。本文用光谱技术对该突变体的光合色素成分进行了鉴定。1 材料和方法 材料为六棱裸大麦“矮杆齐”和由该品种大麦诱变形成的黄绿色突变体1832C(Mb1832C),以及作为对照的缺失Chlb的突变体大麦Chlorina-f2[2]都于3月初播种于实验田中。 每个样品取30g新鲜的叶片,先用自来水后用蒸馏水冲洗干净。把洗净的叶片摊放在干净的纱布上吸干表面水分,剪碎,加入100mL预冷的含有0.4mol/L山梨醇、0.1mol/LTris-HCl(pH7.6)的缓冲液,用组织捣碎机先慢速捣碎1… 相似文献
176.
超结瘤大豆(Glycine m ax (L.) Merr.) nts 382 和不结瘤大豆Nod 49 的叶和根组织水提取物经Sephadex G25 过滤、洗脱,再根据洗脱物对硝酸还原酶(NR)活性的影响可划分为4 个组分(fraction)样品,即nts 382(Nod 49) F1、nts 382(Nod 49) F2、nts 382(Nod 49) F3 和nts 382(Nod 49) F4。其中, nts382 F2 和F4 抑制NR 活性作用在接种USDA110 后明显下降, 但接种的nts 382 F2 却能提高大豆Bragg 的结瘤数达一倍, 而接种的nts 382 F3 和F4 的作用不明显。NR 活性抑制因子不是刺激结瘤的因子, 刺激结瘤的因子主要分布在接种的nts382 F2 部分中。与这一现象相反, Nod 49 F2 和F4 抑制NR活性的作用在接种后更强, 且也抑制大豆nts 382 的结瘤, 其中Nod 49 F4 抑制结瘤的作用基本不能逆转。抑制结瘤因子主要分布在接过种的Nod 49 F4 部分中 相似文献
177.
Xianzhen Li Yunzhan Huang Degui Xu Dongping Xiao Fengxie Jin Peiji Gao 《Biotechnology letters》1996,18(2):205-210
Summary The cellobiose oxidizing enzyme of the newly isolated cellulolytic bacterium Cytophaga sp. LX-7 was produced extracellularly when grown on cellulose or other saccharides, which was previously noted only in fungi. The enzyme could use not only cellobiose, maltose, glucose and other saccharides but also cellulose as substrates, and use dichlorophenol indophenol and oxygen as electron acceptors. 相似文献
178.
Stanislav D. Zakharov Xia Li Taya P. Red'ko Richard A. Dilley 《Journal of bioenergetics and biomembranes》1996,28(6):483-494
The 8-kDa subunit c of theE. coli F0 ATP-synthase proton channel was tested for Ca++ binding activity using a45Ca++ ligand blot assay after transferring the protein from SDS-PAGE gels onto polyvinyl difluoride membranes. The purified subunit c binds45Ca++ strongly with Ca++ binding properties very similar to those of the 8-kDa CF0 subunit III of choloroplast thylakoid membranes. The N-terminal f-Met carbonyl group seems necessary for Ca++ binding capacity, shown by loss of Ca++ binding following removal of the formyl group by mild acid treatment. The dicyclohexylcarbodiimide-reactive Asp-61 is not involved in the Ca++ binding, shown by Ca++ binding being retained in twoE. coli mutants, Asp61Asn and Asp61Gly. The Ca++ binding is pH dependent in both theE. coli and thylakoid 8-kDa proteins, being absent at pH 5.0 and rising to a maximum near pH 9.0. A treatment predicted to increase the Ca++ binding affinity to its F0 binding site (chlorpromazine photoaffinity attachment) caused an inhibition of ATP formation driven by a base-to-acid pH jump in whole cells. Inhibition was not observed when the Ca++ chelator EGTA was present with the cells during the chlorpromazine photoaffinity treatment. An apparent Ca++ binding constant on the site responsible for the UV plus chlorpromazine effect of near 80–100 nM was obtained using an EGTA-Ca++ buffer system to control free Ca++ concentration during the UV plus chlorpromazine treatment. The data are consistent with the notion that Ca++ bound to the periplasimic side of theE. coli F0 proton channel can block H+ entry into the channel. A similar effect occurs in thylakoid membranes, but the Ca++ binding site is on the lumen side of the thylakoid, where Ca++ binding can modulate acid-base jump ATP formation. The Ca++ binding to the F0 and CF0 complexes is consistent with a pH-dependent gating mechanism for control of H+ ion flux across the opening of the H+ channel.This work was supported in part by grants from the Department of Energy and the U.S. Department of Agriculture.On leave from the Institute of Soil Science and Photosynthesis, Russian Academy of Science, Pushchino, Russia. 相似文献
179.
180.
The scl gene product is required for the generation of all hematopoietic lineages in the adult mouse. 总被引:35,自引:4,他引:31 下载免费PDF全文
L Robb N J Elwood A G Elefanty F Kntgen R Li L D Barnett C G Begley 《The EMBO journal》1996,15(16):4123-4129
Homozygosity for a null mutation in the scl gene causes mid-gestational embryonic lethality in the mouse due to failure of development of primitive hematopoiesis. Whilst this observation established the role of the scl gene product in primitive hematopoiesis, the death of the scl null embryos precluded analysis of the role of scl in later hematopoietic development. To address this question, we created embryonic stem cell lines with a homozygous null mutation of the scl gene (scl-/-) and used these lines to derive chimeric mice. Analysis of the chimeric mice demonstrates that the scl-/- embryonic stem cells make a substantial contribution to all non-hematopoietic tissues but do not contribute to any hematopoietic lineage. These observations reveal a crucial role for the scl gene product in definitive hematopoiesis. In addition, in vitro differentiation assays with scl-/- embryonic stem cells showed that the scl gene product was also required for formation of hematopoietic cells in this system. 相似文献