全文获取类型
收费全文 | 3380篇 |
免费 | 265篇 |
国内免费 | 3篇 |
出版年
2024年 | 10篇 |
2023年 | 12篇 |
2022年 | 54篇 |
2021年 | 91篇 |
2020年 | 53篇 |
2019年 | 75篇 |
2018年 | 114篇 |
2017年 | 101篇 |
2016年 | 132篇 |
2015年 | 196篇 |
2014年 | 215篇 |
2013年 | 231篇 |
2012年 | 307篇 |
2011年 | 295篇 |
2010年 | 145篇 |
2009年 | 143篇 |
2008年 | 205篇 |
2007年 | 207篇 |
2006年 | 148篇 |
2005年 | 152篇 |
2004年 | 175篇 |
2003年 | 116篇 |
2002年 | 106篇 |
2001年 | 71篇 |
2000年 | 73篇 |
1999年 | 44篇 |
1998年 | 32篇 |
1997年 | 17篇 |
1996年 | 13篇 |
1995年 | 14篇 |
1994年 | 13篇 |
1993年 | 4篇 |
1992年 | 16篇 |
1991年 | 14篇 |
1990年 | 7篇 |
1989年 | 9篇 |
1988年 | 2篇 |
1987年 | 4篇 |
1986年 | 4篇 |
1984年 | 4篇 |
1983年 | 7篇 |
1982年 | 2篇 |
1980年 | 2篇 |
1977年 | 3篇 |
1972年 | 1篇 |
1971年 | 1篇 |
1967年 | 1篇 |
1965年 | 1篇 |
1960年 | 2篇 |
1959年 | 1篇 |
排序方式: 共有3648条查询结果,搜索用时 15 毫秒
971.
R S English J S Lampel TJ Vanden Boom 《Journal of industrial microbiology & biotechnology》1998,21(4-5):219-224
The introduction of plasmid DNA into germinating spores of an industrially improved strain of Saccharopolyspora erythraea was accomplished by electroporation. Various parameters affecting the efficiency of electroporation were examined. The most
critical factor was the extent of spore germination. Electrocompetence was limited to a 4-h period following the initial emergence
of the germ tube. Electroporation efficiencies as high as 2 × 105 CFU μg−1 of plasmid DNA were obtained using electrocompetent germlings. The optimal field strength was 12–14 kV cm−1 with a pulse duration of 15–20 ms. Electrocompetent germlings were stored at −80°C without a significant decrease in transformation
efficiency. The utility of this protocol was demonstrated by isolating a propionyl-CoA carboxylase mutant through targeted
gene disruption and replacement.
Received 3 April 1998/ Accepted in revised form 28 September 1998 相似文献
972.
A method for the formation and regeneration of protoplasts of several strains of the chestnut blight fungus,Cryphonectria parasitica, is presented. The procedure utillizes cellophane membranes for growth and employs centrifugation for separation of protoplasts from hyphal fragments. Yields averaged 8.04×106 protoplasts per membrane. Regeneration frequencies were 40–50% with a soft-agar overlay. These protoplasts are suitable for use in experiments designed to determine the role of dsRNA in hypovirulence ofC. parasitica. 相似文献
973.
Sequence variations in small-subunit ribosomal RNAs of Hartmannella vermiformis and their phylogenetic implications 总被引:1,自引:0,他引:1
Evidence of associations between free-living amoebas and human disease has
been increasing in recent years. Knowledge about phylogenetic relationships
that may be important for the understanding of pathogenicity in the genera
involved is very limited at present. Consequently, we have begun to study
these relationships and report here on the phylogeny of Hartmannella
vermiformis, a free-living amoeba that can harbor the etiologic agent of
Legionnaires' disease. Our analysis is based on studies of small-subunit
ribosomal RNA genes (srDNA). Nucleotide sequences were determined for
nuclear srDNA from three strains of H. vermiformis isolated from the United
Kingdom, Germany, and the United States. These sequences then were compared
with a sequence previously obtained for a North American isolate by J. H.
Gunderson and M. L. Sogin. The four genes are 1,840 bp long, with an
average GC content of 49.6%. Sequence differences among the strains range
are 0.38%-0.76%. Variation occurs at 19 positions and includes 2
single-base indels plus 14 monotypic and 3 ditypic single-base
substitutions. Variation is limited to eight helix/loop structures
according to a current model for srRNA secondary structure. Parsimony,
distance, and bootstrap analyses used to examine phylogenetic relationships
between the srDNA sequences of H. vermiformis and other eukaryotes
indicated that Hartmannella sequences were most closely related to those of
Acanthamoeba and the alga Cryptomonas. All ditypic sites were consistent
with a separation between European and North American strains of
Hartmannella, but results of other tests of this relationship were
statistically inconclusive.
相似文献
974.
975.
Ouabain-insensitive salt and water movements in duck red cells. I. Kinetics of cation transport under hypertonic conditions 总被引:10,自引:6,他引:4
下载免费PDF全文
![点击此处可从《The Journal of general physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Duck red cells in hypertonic media experience rapid osmotic shrinkage followed by gradual reswelling back toward their original volume. This uptake of salt and water is self limiting and demands a specific ionic composition of the external solution. Although ouabain (10(-4)M) alters the pattern of cation accumulation from predominantly potassium to sodium, it does not affect the rate of the reaction, or the total amount of salt or water taken up. To study the response without the complications of active Na-K transport, ouabain was added to most incubations. All water accumulated by the cells can be accounted for by net salt uptake. Specific external cation requirements for reswelling include: sufficient sodium (more than 23 mM), and elevated potassium (more than 7 mM). In the absence of external potassium cells lose potassium without gaining sodium and continue to shrink instead of reswelling. Adding rubidium to the potassium- free solution promotes an even greater loss of cell potassium, yet causes swelling due to a net uptake of sodium and rubidium followed by chloride. The diuretic furosemide (10(-3)M) inhibits net sodium uptake which depends on potassium (or rubidium), as well as inhibits net sodium uptake which depends on sodium. As a result, cell volume is stabilized in the presence of this drug by inhibition of shrinkage, at low, and of swelling at high external potassium. The response has a high apparent energy of activation (15-20 kcal/mol). We propose that net salt and water movements in hypertonic solutions containing ouabain are mediated by direct coupling or cis-interaction, between sodium and potassium so that the uphill movement of one is driven by the downhill movement of the other in the same direction. 相似文献
976.
Detection of ischemia-reperfusion cardiac injury by cardiac muscle chemiluminescence 总被引:2,自引:0,他引:2
Kailash Prasad Paul Lee Subrahmanyam V. Mantha Jawahar Kalra Marion Prasad Jang B. Gupta 《Molecular and cellular biochemistry》1992,115(1):49-58
Various methods have been used in the past to assess the implication of oxygen free radicals (OFR) in ischemia-reperfusion-induced cardiac injury. Luminol-enhanced tert-butyl-initiated chemiluminescence in cardiac tissue reflects oxidative stress and is a very sensitive method. It was used to elucidate the role of OFR in cardiac injury due to ischemia and reperfusion. Studies were conducted on perfused isolated rabbit hearts in three groups (n = 8 in each): I, control; II, submitted to global ischemia for 30 min; III, submitted to ischemia for 30 min followed by reperfusion for 60 min. The heart tissue was then assayed for chemiluminescence (CL); content of malondialdehyde (MDA), an indicator of OFR-induced cardiac injury; and activity of tissue levels of antioxidants [superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px)].The control values for left and right ventricular CL and malondialdehyde were 81.1 ± 15.4 (S.E.) and 182.4 ± 50.3 (S.E.), mv-min-mg protein–1; and 0.024 ± 0.006 (S.E.) and 0.324 ± 0.005 (S.E.) nmoles-mg protein–1 respectively. Ischemia produced an increase in the cardiac CL (3.3 to 4.4 fold) and MDA content (2 to 2.6 fold). Reperfusion following ischemia also produced similar changes in CL and MDA content. The control values for activity of left ventricular SOD, catalase, and GSH-Px were 45.77 ± 1.73 (S.E.) U-mg protein–1 5.35 ± 0.51 (S.E.) K-10–3-sec–1-mg protein–1, and 77.50 ± 7.70 (S.E.) nmoles NADPH-min–1-mg protein–1 respectively. Activities of SOD and catalase decreased during ischemia but were similar to control values in ischemic-reperfused hearts. The GSH-Px activity of left ventricle was unaffected by ischemia, and ischemia-reperfusion. GSH-Px activity of the right ventricle increased with ischemia, and ischemic-reperfusion.These results indicate that cardiac tissue chemiluminescence would be a useful and sensitive tool for the detection of oxygen free radical-induced cardiac injury. 相似文献
977.
Sung R. Min Seung G. Yang Jang R. Liu Pil S. Choi Woong Y. Soh 《Plant cell reports》1992,10(12):621-623
Culture conditions for high frequency somatic embryogenesis and plant regeneration from cotyledonary explants of Codonopsis lanceolata are described. The maximum induction frequency of somatic embryos from cotyledonary explants was 80% on Murashige and Skoog (MS) medium containing 6% sucrose with 1 mg/l 2,4-dichlorophenoxyacetic acid and 10% coconut water. Upon transfer onto MS basal medium containing 3% sucrose, most somatic embryos developed into plantlets.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellin a3
- MS
Murashige and Skoog 相似文献
978.
Hyeon-Ok Jin Sung-Eun Hong Ji-Young Kim Se-Kyeong Jang In-Chul Park 《Cell death & disease》2021,12(12)
Amino acid availability is sensed by various signaling molecules, including general control nonderepressible 2 (GCN2) and mechanistic target of rapamycin complex 1 (mTORC1). However, it is unclear how these sensors are associated with cancer cell survival under low amino acid availability. In the present study, we investigated AKT activation in non-small cell lung cancer (NSCLC) cells deprived of each one of 20 amino acids. Among the 20 amino acids, deprivation of glutamine, arginine, methionine, and lysine induced AKT activation. AKT activation was induced by GCN2/ATF4/REDD1 axis-mediated mTORC2 activation under amino acid deprivation. In CRISPR-Cas9-mediated REDD1-knockout cells, AKT activation was not induced by amino acid deprivation, indicating that REDD1 plays a major role in AKT activation under amino acid deprivation. Knockout of REDD1 sensitized cells cultured under glutamine deprivation conditions to radiotherapy. Taken together, GCN2/ATF4/REDD1 axis induced by amino acid deprivation promotes cell survival signal, which might be a potential target for cancer therapy.Subject terms: Cancer metabolism, Cell death 相似文献
979.
Kyoo Heo Jae-Woo Lee Yongdae Jang Sohee Kwon Jaehun Lee Chaok Seok Nam-Chul Ha Yeong-Jae Seok 《The Journal of biological chemistry》2022,298(3)
The bacterial second messenger bis-(3′-5′)-cyclic diguanylate monophosphate (c-di-GMP) controls various cellular processes, including motility, toxin production, and biofilm formation. c-di-GMP is enzymatically synthesized by GGDEF domain–containing diguanylate cyclases and degraded by HD-GYP domain–containing phosphodiesterases (PDEs) to 2 GMP or by EAL domain–containing PDE-As to 5ʹ-phosphoguanylyl-(3ʹ,5ʹ)-guanosine (pGpG). Since excess pGpG feedback inhibits PDE-A activity and thereby can lead to the uncontrolled accumulation of c-di-GMP, a PDE that degrades pGpG to 2 GMP (PDE-B) has been presumed to exist. To date, the only enzyme known to hydrolyze pGpG is oligoribonuclease Orn, which degrades all kinds of oligoribonucleotides. Here, we identified a pGpG-specific PDE, which we named PggH, using biochemical approaches in the gram-negative bacteria Vibrio cholerae. Biochemical experiments revealed that PggH exhibited specific PDE activity only toward pGpG, thus differing from the previously reported Orn. Furthermore, the high-resolution structure of PggH revealed the basis for its PDE activity and narrow substrate specificity. Finally, we propose that PggH could modulate the activities of PDE-As and the intracellular concentration of c-di-GMP, resulting in phenotypic changes including in biofilm formation. 相似文献
980.
J J Sekelsky K S McKim L Messina R L French W D Hurley T Arbel G M Chin B Deneen S J Force K L Hari J K Jang A C Lauren?on L D Madden H J Matthies D B Milliken S L Page A D Ring S M Wayson C C Zimmerman R S Hawley 《Genetics》1999,152(2):529-542
The segregation of homologous chromosomes from one another is the essence of meiosis. In many organisms, accurate segregation is ensured by the formation of chiasmata resulting from crossing over. Drosophila melanogaster females use this type of recombination-based system, but they also have mechanisms for segregating achiasmate chromosomes with high fidelity. We describe a P-element mutagenesis and screen in a sensitized genetic background to detect mutations that impair meiotic chromosome pairing, recombination, or segregation. Our screen identified two new recombination-deficient mutations: mei-P22, which fully eliminates meiotic recombination, and mei-P26, which decreases meiotic exchange by 70% in a polar fashion. We also recovered an unusual allele of the ncd gene, whose wild-type product is required for proper structure and function of the meiotic spindle. However, the screen yielded primarily mutants specifically defective in the segregation of achiasmate chromosomes. Although most of these are alleles of previously undescribed genes, five were in the known genes alphaTubulin67C, CycE, push, and Trl. The five mutations in known genes produce novel phenotypes for those genes. 相似文献