全文获取类型
收费全文 | 2405篇 |
免费 | 158篇 |
国内免费 | 3篇 |
出版年
2023年 | 15篇 |
2022年 | 27篇 |
2021年 | 103篇 |
2020年 | 33篇 |
2019年 | 44篇 |
2018年 | 80篇 |
2017年 | 56篇 |
2016年 | 90篇 |
2015年 | 102篇 |
2014年 | 152篇 |
2013年 | 193篇 |
2012年 | 236篇 |
2011年 | 246篇 |
2010年 | 141篇 |
2009年 | 130篇 |
2008年 | 153篇 |
2007年 | 131篇 |
2006年 | 121篇 |
2005年 | 86篇 |
2004年 | 94篇 |
2003年 | 65篇 |
2002年 | 65篇 |
2001年 | 20篇 |
2000年 | 18篇 |
1999年 | 11篇 |
1998年 | 5篇 |
1997年 | 5篇 |
1996年 | 5篇 |
1995年 | 11篇 |
1994年 | 4篇 |
1993年 | 8篇 |
1992年 | 10篇 |
1991年 | 10篇 |
1990年 | 8篇 |
1989年 | 5篇 |
1988年 | 9篇 |
1987年 | 10篇 |
1986年 | 7篇 |
1985年 | 9篇 |
1983年 | 7篇 |
1982年 | 2篇 |
1979年 | 5篇 |
1978年 | 5篇 |
1977年 | 3篇 |
1976年 | 3篇 |
1975年 | 3篇 |
1974年 | 3篇 |
1973年 | 2篇 |
1970年 | 3篇 |
1962年 | 2篇 |
排序方式: 共有2566条查询结果,搜索用时 15 毫秒
11.
Expression and biochemical characterization of human immunodeficiency virus type 1 nef gene product. 总被引:11,自引:6,他引:5
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)
J Kaminchik N Bashan D Pinchasi B Amit N Sarver M I Johnston M Fischer Z Yavin M Gorecki A Panet 《Journal of virology》1990,64(7):3447-3454
nef genes from human immunodeficiency virus type 1 isolates BH10 and LAV1 (lymphadenopathy-associated virus type 1) were expressed in Escherichia coli under the deo operon promoter. The two proteins found in the soluble compartment of the bacterial lysate were purified by ion-exchange column chromatography to apparent homogeneity. Determination of the amino-terminal sequence revealed glycine as the first amino acid in the Nef protein, indicating removal of the initiator methionine during expression in E. coli. Under native conditions, the recombinant Nef protein is a monomer of 23 kilodaltons. In denaturing polyacrylamide gels, however, BH10 and LAV1 Nef proteins migrate as 28 and 26 kilodaltons, respectively. GTP binding and GTPase activity were monitored during Nef protein purification. These activities did not copurify with the recombinant Nef protein from either the BH10 or the LAV1 isolate. Purified recombinant BH10 Nef protein was used as an immunogen to elicit mouse monoclonal antibodies. A series of monoclonal antibodies were obtained which reacted with sequences at either the amino or carboxy terminus of Nef. In addition, a conformational epitope reacting with native BH10, but not LAV1, Nef was isolated. 相似文献
12.
Amit Banerjee Peter C. Weber Sunil Palchaudhuri 《Molecular & general genetics : MGG》1990,220(2):320-324
Summary We have isolated a deletion mutation and a point mutation in the copB gene of the replication region Repl of the IncFI plasmid Co1V2-K94. Subsequently, this copB gene with and without point mutation was cloned and sequenced, and the point mutation was mapped in the coding region of copB with a change of one amino acid from arginine to serine. Furthermore, this copB mutant had an approximately 10-fold increase in copy number. The CopB-phenotype of Co1V2-K94 could be complemented in trans by the copB gene of coresident IncFII plasmids such as R1 and R538, but not R100, suggesting that ColV2-K94 and R1 or R538 contain the same copB allele. 相似文献
13.
Raman Kapur Mohammed Saleem Bryan L. Harvey Adrian J. Cutler 《In vitro cellular & developmental biology. Plant》1993,29(4):200-206
Summary Barley leaf blade protoplasts accumulate malonaldehyde, a product of lipid peroxidation, during culture. In addition, glutathione
levels fall after protoplast isolation and the proportion of glutathione in the oxidized state rises. These data indicate
oxidative stress after protoplast isolation and during culture. The cause of this phenomenon is revealed by data showing that
the activities of enzymes associated with antioxidative processes including glutathione reductase and ascorbate peroxidase
decrease after barley protoplast isolation. In contrast, protoplasts isolated from suspension cultured cells of bromegrass
and soybean exhibit little evidence for oxidative stress and increased activities of glutathione reductase and ascorbate peroxidase.
We suggest that an antioxidative response is associated with mitosis and colony formation from protoplasts, as exhibited by
bromegrass and soybean. Conversely, failure of an antioxidative response is associated with low viability and absence of mitosis,
as in barley. Increased viability of barley leaf protoplasts cultured on feeder layer cells is correlated with increased glutathione
content and higher glutathione reductase activity. 相似文献
14.
Sharma Shivani Kumar Amit Dhakte Priyanka Raturi Gaurav Vishwakarma Gautam Barbadikar Kalyani M. Das B. K. Shivaraj S. M. Sonah Humira Deshmukh Rupesh 《Journal of Plant Growth Regulation》2023,42(1):46-59
Journal of Plant Growth Regulation - Crop improvement in light of the rapidly changing climate and the increasing human population continues to be one of the primary concerns for researchers across... 相似文献
15.
Effects of anatomic variability on blood flow and pressure gradients in the pulmonary capillaries 总被引:1,自引:0,他引:1
Dhadwal Amit; Wiggs Barry; Doerschuk Claire M.; Kamm Roger D. 《Journal of applied physiology》1997,83(5):1711-1720
Dhadwal, Amit, Barry Wiggs, Claire M. Doerschuk, and RogerD. Kamm. Effects of anatomic variability on blood flow and pressure gradients in the pulmonary capillaries. J. Appl. Physiol. 83(5): 1711-1720, 1997.Atheoretical model is developed to simulate the flow of blood throughthe capillary network in a single alveolar septum. The objective is tostudy the influence of random variability in capillary dimension andcompliance on flow patterns and pressures within the network. Thecapillary bed is represented as an interconnected rectangular grid ofcapillary segments and junctions; blood flow is produced by applying apressure gradient across the network. Preferred flow channels are shownto be a natural consequence of random anatomic variability, the effectof which is accentuated at low transcapillary pressures. Thedistribution of pressure drops across single capillary segments widenswith increasing network variability and decreasing capillary transmuralpressure. Blockage of one capillary segment causes the pressure dropacross that segment to increase by 60%, but the increase falls to<10% at a distance of three segments. The factors that causenonuniform capillary blood flow through the capillary network arediscussed. 相似文献
16.
David A. Watson Vivek Kapur Daniel M. Musher James W. Jacobson James M. Musser 《Current microbiology》1995,31(4):251-259
This paper reports the cloning and sequencing of a region of DNA from Streptococcus pneumoniae serotype 3 surrounding transposon Tn916, insertion of which was previously shown to result in lack of expression of the extracellular capsule. Sequence analysis revealed that the transposon inserted into a consensus insertion site 71 bp from the 5 end of the cloned fragment. Within the clone, 3 downstream regions from two different pneumococcal lytA genes were identified, as well as a putative 194 AA open reading frame (ORF1). Moreover, two copies of the repeat element BOX, oriented in opposite directions, were located immediately 3 of orf1. Within the region bounded by the first pair of internal sequencing primers, analysis revealed that the fragment amplified by PCR was always of the same size. Moreover, Southern blotting showed that for all serotypes examined to date, homology exists with the cloned fragment. These results indicate that this region of the chromosome is highly conserved and, taken together with other independently derived data, suggest that interruptions or deletions within this DNA lead to unencapsulation. 相似文献
17.
In vivo and in vitro activities of nitrate reductase were assayedin Crotalaria juncea pollen suspension cultures. This enzymewas found to be substrate-inducible and enhanced activity wasobserved when it was extracted with cysteine buffer or incubatedwith NADH (0.6 mM) at 25?C or when the germinated pollen grainswere treated with red light for 10 min. Enzymes of ammonia assimilation,glutamate dehydrogenase and glutamate synthetase, and also thepentose phosphate-shunt enzyme, glucose-6-phosphate dehydrogenase,which catalyzes the step that provides reducing power to thesystem, are described. (Received October 20, 1977; ) 相似文献
18.
R S Sharma S Mokkapati M Rajalakshmi M M Kapur 《Indian journal of experimental biology》1992,30(11):1079-1083
Attempts were made to validate RIA for prolactin (PRL), LH and FSH in semen from normospermic, oligospermic and azoospermic subjects. The RIA used to measure PRL and LH in semen fulfilled the criteria of reliability, whereas low levels of FSH in semen precluded the validation of FSH assay in semen. Semen levels of PRL and LH were significantly (P < 0.05) higher than serum levels in all groups of subjects investigated. Semen levels of FSH in azoospermic men after vasoepididymostomy (VEA), were significantly (P < 0.05) higher compared to azoospermic men prior to surgery. Serum levels of PRL were significantly higher (P < 0.05) in normospermic men compared to oligospermic and azoospermic men prior to and after surgery. Semen levels of PRL in normospermic men were comparable with oligospermic and azoospermic subjects prior to and after surgery. Serum levels of LH in oligospermic and azoospermic men who did not undergo surgery and in men reporting oligospermia after VEA were comparable to normospermic subjects but in men showing azoospermia post surgically, serum LH levels were significantly (P < 0.005) elevated. Semen levels of LH in men reporting azoospermia before surgery and in subjects showing oligospermia or azoospermia post surgically were significantly lower (P < 0.05) compared to men with normal sperm count. Serum levels of FSH were significantly elevated (P < 0.05) compared to semen levels in oligospermic men prior to surgery but this increase was not seen in post VEA subjects. These results were discussed. 相似文献
19.
Ramesh N. Patel Mark Liu Amit Banerjee Laszlo J. Szarka 《Applied microbiology and biotechnology》1992,37(2):180-183
Summary A key chiral intermediate lactol(3)[3aS (3a,4,7,7a)]-hexahydro-4,7-epoxy-isobenzofuran-1 (3H)-one was prepared for the total synthesis of a new thromboxane antagonist. The stereoselective hydrolysis of (exo,exo)-7-oxabicyclo[2.2.1]heptane-2,3-dimethanol, diacetate ester (1) to the corresponding chiral monoacetate ester (2) was carried out with lipases, among which Amano P-30 lipase from Pseudomonas sp. was most effective since it gave the desired enantiomer of monoacetate ester. A yield of 75 mol% and optical purity of >99% was obtained when the reaction was conducted in a biphasic system with 10% toluene at 5 g/l of the substrate. Lipase P-30 was immobilized on Accurel polypropylene (PP) and the immobilized enzyme was reused (five cycles) without loss of enzyme activity, productivity or optical purity. The reaction process was scaled-up to 80 1 (400 g substrate) and monoacetate (2) was isolated in 80 mol% yield with 99.3% optical purity as determined by chiral HPLC and nuclear magnetic resonance (NMR) analysis. A gas chromatography of 99.5% and specific rotation, []D of -7.6° was obtained. The chiral monoacetate ester (2) was oxidized to its corresponding aldehyde and subsequently hydrolyzed to give lactol (3). 相似文献
20.
The 5' flanking region from the human dopamine beta-hydroxylase gene directs expression of bacterial beta-galactosidase reporter genes to a subset of adult neurons and adrenal chromaffin cells of transgenic mice. In this paper, we examine the spatial and temporal patterns of expression of these transgenes during embryogenesis. Expression begins at embryonic day 9 in the developing central and peripheral nervous systems and persists in cell populations in which expression is observed in adult transgenic mice. However, transient embryonic expression occurs in presumptive neuroblasts in developing sensory ganglia and ventrolateral neural tube that are destined to synthesize neurotransmitters other than catecholamines. These observations support the concept that some cells fated to become "non-catecholaminergic" neurons exhibit transient catecholaminergic features during their differentiation. 相似文献