全文获取类型
收费全文 | 71100篇 |
免费 | 5047篇 |
国内免费 | 3227篇 |
出版年
2024年 | 126篇 |
2023年 | 1027篇 |
2022年 | 1682篇 |
2021年 | 2341篇 |
2020年 | 2184篇 |
2019年 | 2464篇 |
2018年 | 2457篇 |
2017年 | 1761篇 |
2016年 | 1746篇 |
2015年 | 2254篇 |
2014年 | 4249篇 |
2013年 | 5295篇 |
2012年 | 3190篇 |
2011年 | 4306篇 |
2010年 | 3309篇 |
2009年 | 3681篇 |
2008年 | 3756篇 |
2007年 | 3821篇 |
2006年 | 3399篇 |
2005年 | 3039篇 |
2004年 | 2704篇 |
2003年 | 2270篇 |
2002年 | 2027篇 |
2001年 | 1395篇 |
2000年 | 1179篇 |
1999年 | 1222篇 |
1998年 | 1124篇 |
1997年 | 981篇 |
1996年 | 924篇 |
1995年 | 846篇 |
1994年 | 768篇 |
1993年 | 709篇 |
1992年 | 611篇 |
1991年 | 577篇 |
1990年 | 446篇 |
1989年 | 411篇 |
1988年 | 376篇 |
1987年 | 353篇 |
1986年 | 315篇 |
1985年 | 435篇 |
1984年 | 620篇 |
1983年 | 471篇 |
1982年 | 517篇 |
1981年 | 360篇 |
1980年 | 361篇 |
1979年 | 301篇 |
1978年 | 221篇 |
1977年 | 172篇 |
1976年 | 143篇 |
1975年 | 131篇 |
排序方式: 共有10000条查询结果,搜索用时 62 毫秒
891.
Richard A. Bafford Robert W. Seagull Si-Yin Chung David F. Millie 《Journal of phycology》1993,29(1):91-95
The monoterpene derivative, 2-methylisoborneol (MIB), is produced by many blue-green algae and often is responsible for the “musty” taste/odor in aquaculturally raised finfish and potable water supplies. Although previous researchers have suggested that taste/odor metabolites are partitioned among cell constituents and that coregulation with pigment biosynthesis occurs, no structural evidence for these hypotheses exists. MIB was localized in cells of Oscillatoria limosa (Roth) Agardh at the ultrastructural level using standard gold-labeled antibody techniques. There was no apparent relationship between age of the cells and MIB synthesis; cells that were and were not undergoing active division had similar amounts of MIB label. There was no consistent partitioning of MIB label within cells. However, occasionally, specific label was observed along photosynthetic lamellae, suggesting a potential linkage of MIB synthesis and/or binding to the pigment systems. 相似文献
892.
Gregory M. L. Patterson Kathleen K. Baker Cynthia L. Baldwin Christine M. Bolis Faith R. Caplan Linda K. Larsen Ira A. Levine Richard E. Moore E. Moore Carrie S. Nelson Kathryn D. Tschappat Grace D. Tuang Michael R. Boyd John H. Cardellina Ralph P. Collins Kirk R. Gustafson Kenneth M. Snader Owen S. Weislow Ralph A. Lewin 《Journal of phycology》1993,29(1):125-130
Lipophilic and hydrophilic extracts from approximately 600 strains of cultured cyanophytes, representing some 300 species, were examined for antiviral activity against three pathogenic viruses. Approximately 10% of the cultures produced substances that caused significant reduction in cytopathic effect normally associated with viral infection. The screening program identified the order Chroococcales as commonly producing antiviral agents. 相似文献
893.
Abstract: The present study reports the ion dependency of 2β-carbomethoxy-3β-(4-fluorophenyl)[3 H]tropane ([3 H]- CFT) binding to the dopamine transporter in the rat striaturn. The results indicate that [3 H]CFT binding to synaptosomal P2 membranes requires low concentrations of Na+ (peak binding between 20 and 50 m M Na+ ), is stimulated by phosphate anion or l- , but is unaffected or only slightly affected by F- , Cl- , Br- , NO3 - , or SO4 2- , Concentrations of Na+ of >50 m M become inhibitory except in the presence of l- , which shifts peak binding levels toward higher Na+ concentrations and also elevates the peak binding level. K+ strongly decreased [3 H]CFT binding with a shallow inhibition curve, and Na+ could not overcome this effect. Saturation analysis of [3 H]CFT binding revealed a single binding site changing its affinity for CFT depending on the concentration of sodium phosphate buffer (6, 10, 30, 50, 130, or 200 m M ; 1 mM plus 49 mM NaCIversus 10 m M plus 40 m M NaCI; or 1 mM plus 129 m M Nal versus 10 m M plus 120 m M Nal). No differences were observed in the density of CFT binding sites between any of the conditions examined. 相似文献
894.
Commercially available lactase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) enzymes produced from Kluyveromyces fragilis and Kluyveromyces lactis were accessed as catalysts for use in the production of beta-galactopyranosides of various alcohols using lactose as galactosyl donor. The yield of galactoside was enhanced by using the highest practical concentrations of both lactose and alcohol acceptor. The concentrations and thus yield, were limited by the solubility of the substrates. The increase in galactoside yield with increasing lactose concentration appeared to be specific to the lactose substrate and not due to water activity alterations, because addition of maltose to a fixed concentration of lactose had no effect. During the course of the reaction, the yield of galactoside peaked after around 70% to 80% of the lactose was consumed, due to hydrolysis of the product by the enzyme. A wide variety of compounds with primary or secondary hydroxyl groups could act as acceptors, the essential requirement being at least some water solubility. Addition of organic cosolvents had little effect on galactoside yield except when it increased the water solubility of sparingly soluble alcohols. Some galactosides were synthesized on a gram scale to determine practical product recoveries and improve purification methods for large-scale synthesis. Initial purification by hydrophobic chromatography (for galactosides of hydrophobic alcohols) or strong anion-exchange chromatography (for galactosides of hydrophilic alcohols) separated galactosides, galactobiosides, and higher oligomers from reducing sugars. A facile separation of the galactoside and galactobioside could then be effected by flash chromatography on silica gel. (c) 1993 John Wiley & Sons, Inc. 相似文献
895.
Park YS Shiba S Lijima S Kobayashi T Hishinuma F 《Biotechnology and bioengineering》1993,41(9):854-861
Cloned gene expression in recombinant Saccharomyces cerevisiae 20B-12 containing three different plasmids was compared in batch and fed-batch cultures. The plasmids pNA3, pNA7, and pNA9 contain the alpha-amylase gene under the control of SUC2, PGK, and GAL7 Promoters, respectively. The synthesis of alpha-amylase was therefore induced by low glucose concentration for the SUC2 and PGK promoters, and by galactose for GAL7 promoter. The specific cell growth rates were similar among cells harboring the three different plasmids; they decreased from 0.35 to 0.38 h(-1) during the cell growth phase to 0.03 to 0.06h(-1) during the production phase. The secretory alpha-amylase activity of cells harboring plasmid pNA7 was 129 U/mL in fed-batch culture, which was 1.4 and 2 times as high as the activities of cells harboring plasmids pNA3 and pNA9, respectively. The secretion ratios (amount of extracellular alpha-amylase activity/amounts of total alpha-amylase activity) of cells harboring plasmids pNA3, pNA7, and pNA9 were 91.4%, 94.5%, and 95.3%, respectively. (c) 1993 Wiley & Sons, Inc. 相似文献
896.
A rapid two-step procedure has been developed for the purification of Despro(2)-Val15-Leu17-aprotinin from the culture supernatant of a recombinant yeast by affinity and ion-exchange chromatography. DesPro(2)-Val15-Leu17-aprotinin was purified to homogeneity, as demonstrated by dodecylsulfate gel electrophoresis and analysis of the N-terminal amino acid sequence. (c) 1993 John Wiley & Sons, Inc. 相似文献
897.
Differences in plasmid retention and expression are studied in both suspended and biofilm cultures of Escherichia coli DH5alpha(PMJR1750). An alternative mathematical model is proposed which allows the determination of plasmid loss probability in both suspended batch and continuously fed biofilm cultures. In our experiments, the average probability of plasmid loss of E. coli DH5alpha(pMJR1750) is 0.0022 in batch culture in the absence of antibiotic selection pressure and inducer. Under the induction of 0.17 MM IPTG, the maximum growth rate of plasmid-bearing cells in suspended batch culture dropped from 0.45 h(-1) to 0.35 h(-1) and the beta-galactosidase concentration reached an experimental maximum of 0.32. pg/cell 4 hours after the initiation of induction. At both 0.34 and 0.51 mM IPTG, growth rates in batch cultures decreased to 0.16 h(-1), about 36% of that without IPTG, and the beta-galactosidase concentration reached an experimental maximum of 0.47 pg/cell 3 hours after induction.In biofilm cultures, both plasmid-bearing and plasmid-free cells in increase with time reaching a plateau after 96 hours n the absence of both the inducer and any antibiotic selection pressure. Average probability of plasmid loss for biofilm-bound E. coli DH5beta(pMJR1750) population was 0.017 without antibiotic selection. Once the inducer IPTG was added, the concentration of plasmid-bearing cells in biofilm dropped dramatically while plasmid-free cell numbers maintained unaffected. The beta-galactosidase concentration reached a maximum in all biofilm experiments 24 hours after induction; they were 0.08, 0.1, and 0.12 pg/cel under 0.17, 0.34, and 0.51 mM IPTG, respectively. (c) 1993 John Wiley & Sons, Inc. 相似文献
898.
Extractive separation of penicillin G by facilitated transport via carrier supported liquid membranes 总被引:2,自引:0,他引:2
The facilitated transport of penicillin G (Pen G), through a supported liquid membrane with Amberlite LA-2 dissolved in 1-decanol, supported on a microporous polypropylene membrane, were studied. The distribution coefficient was obtained from a batch extraction experiment. The effects of flow rate, carrier concentration, initial concentration of Pen G, and the pH of feed and stripping phases on the transport rate of Pen G through the supported liquid membrane were also investigated. The results are in agreement with theoretical predictions, and it is demonstrated that the transport of Pen G through the supported liquid membrane is controlled simultaneously by mass transfer across both aqueous and liquid membranes. (c) 1993 John Wiley & Sons, Inc. 相似文献
899.
Recombinant mammalian cultures for heterologous gene expression typically involve cells traversing the cell cycle. Studies were conducted to characterize rates of accumulation of intracellular foreign protein in single cells during the cell cycle of Chinese hamster ovary (CHO) cells transfected with an expression vector containing the gene for dihydrofolate reductase (dhfr) and the lacZ gene for bacterial beta-galactosidase (a nonsecreated protein). The lacZ gene was under the control of the constitutive cytomegalovirus promoter. These normally attachment-grown cells were adapted to suspension culture in 10(-7) M methotrexate, and a dual-laser flow cytometer was used to simultaneously determine the DNA and foreign protein (beta-galactosidase) content of single living cells. Expression of beta-galactosidase as a function of cell cycle phase was evaluated for cells in the exponential growth phase, early plateau phase, and inhibited traverse of the cell cycle during exponential growth. The results showed that the beta-galactosidase production rate is higher in the S phase than that in the G1 or G2/M phases. Also, when cell cycle progression was stopped at the S phase by addition of aphidicolin, beta-galactosidase content in single cells was higher than that in exponential phase or plateau phase cells and increased with increasing culture time. Although the cells did not continue to divide after aphidicolin addition, the production of beta-galactosidase per unit volume of culture was very similar to that in normal exponential growth. (c) 1993 John Wiley & Sons, Inc. 相似文献
900.
The stable continuous overproduction of a plasmidencoded protein, beta-lactamase, for at least 50 days by Escherichia coli K-12, RB791(pKN), with release into the culture medium has been demonstrated in two-stage chemostats. The second-stage culture was continuously induced with 0.1 mM IPTG. Continuous expression of beta-lactamase could not be sustained with this strain in a single-stage chemostat because of cell death and selection for lac(-1) cells. beta-Lactamase production in the second stage was sensitive to the second-stage dilution rate and the distribution of the limiting substrate (i.e., glucose) between the first and second stages. The fraction of viable, excreting cells and the average copy number in the induced culture was measurably higher under those conditions of dilution rate and substrate distribution which yielded high beta-lactamase levels. The best operating conditions found at 20 degrees C were a first-stage dilution rate of 0.12 h(-1), a second-stage dilution rate of 0.03 h(-1), and equal glucose feed supplied to each stage. Enzymatically active beta-lactamase was produced at a level of 25% of total cellular protein with 90% excretion yielding 300 mg beta-lactamase/L that was 50% pure at an OD(600) < 6. (c) 1993 Wiley & Sons, Inc. 相似文献