全文获取类型
收费全文 | 12392篇 |
免费 | 1085篇 |
国内免费 | 3篇 |
专业分类
13480篇 |
出版年
2023年 | 43篇 |
2022年 | 128篇 |
2021年 | 253篇 |
2020年 | 147篇 |
2019年 | 169篇 |
2018年 | 231篇 |
2017年 | 182篇 |
2016年 | 317篇 |
2015年 | 577篇 |
2014年 | 568篇 |
2013年 | 729篇 |
2012年 | 992篇 |
2011年 | 1038篇 |
2010年 | 633篇 |
2009年 | 539篇 |
2008年 | 860篇 |
2007年 | 860篇 |
2006年 | 769篇 |
2005年 | 718篇 |
2004年 | 764篇 |
2003年 | 632篇 |
2002年 | 647篇 |
2001年 | 109篇 |
2000年 | 68篇 |
1999年 | 106篇 |
1998年 | 164篇 |
1997年 | 115篇 |
1996年 | 110篇 |
1995年 | 97篇 |
1994年 | 84篇 |
1993年 | 76篇 |
1992年 | 57篇 |
1991年 | 65篇 |
1990年 | 42篇 |
1989年 | 41篇 |
1988年 | 43篇 |
1987年 | 39篇 |
1986年 | 52篇 |
1985年 | 42篇 |
1984年 | 44篇 |
1983年 | 45篇 |
1982年 | 40篇 |
1981年 | 37篇 |
1980年 | 35篇 |
1979年 | 25篇 |
1978年 | 23篇 |
1977年 | 19篇 |
1976年 | 24篇 |
1974年 | 16篇 |
1973年 | 13篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
21.
Timothy J. Strabala Sebastian Y. Bednarek Gregory Bertoni Richard M. Amasino 《Molecular & general genetics : MGG》1989,216(2-3):388-394
Summary A 1.9 kb clone of the T-DNA region of the Agrobacterium tumefaciens Ti plasmid Bo542 which exhibited homology to the isopentenyl transferase (ipt) locus of pTiA6 was identified by low stringency DNA hybridization. Introduction of this segment of pTiBo542 DNA into cells of Nicotiana tabacum or N. glauca caused tumor formation in vivo, and allowed hormone independent growth in vitro. Furthermore, this DNA segment complemented ipt mutant strains of A. tumefaciens, restoring their ability to cause tumors on Kalanchöe leaves and tomato stems. The complete DNA sequence of this segment has been determined, revealing an open reading frame homologous to other known Agrobacterium ipt genes. 相似文献
22.
An important component of computer programs for determining the solution conformation of proteins and other flexible molecules
from nuclear magnetic resonance data are the so-called “bound smoothing algorithms”, which compute lower and upper limits
on the values of all the interatomic distances from the relatively sparse set which can usually be measured experimentally.
To date, the only methods efficient enough for use in large problems take account of only the triangle inequality, but an
appreciable improvement in the precision of the limits is possible if the algebraic relations between the distances among
each quadruple of atoms are also considered. The goal of this paper is to use a recently improved algorithm for computing
these “tetrangle inequality limits” to determine just how much improvement really is possible, given the types of experimental
data that are usually available. 相似文献
23.
Timothy Troy 《Dialectical Anthropology》1990,15(1):74-81
Timothy Troy is a Librarian at the Center for Creative Photography, University of Arizona. 相似文献
24.
Summary The complete physical map of the mitochondrial genome of the Owen cytoplasm of sugar beet has been determined from overlapping cosmid clones. The genome is 386 kb in size and has a multicircular organisation generated by homologous recombination across repeated DNA elements. The location of the rRNA genes and several polypeptide genes has been determined. In addition the mitochondrial genome was found to contain a sequence of chloroplast DNA including part of the 16 S rRNA gene. 相似文献
25.
Mahan James R.; Sherman Timothy D.; Funkhouser Edward A. 《Plant & cell physiology》1988,29(4):735-737
The thermal dependence of two of the reactions catalyzed bythe nitrate reductase from Chlorella vulgaris was determined.The activation energies for NADH:nitrate oxidoreductase (EC1.6.6.1
[EC]
) and NADH:Cytochrome c oxidoreductase (EC 1.6.99.3
[EC]
)are 42.1 kJ?mol1 and 21.5 kJ?mol1, respectively.Since the thermal dependency of the two enzymes is different,ratios of the activities will vary with temperature. The importanceof both rigorous thermal control during nitrate reductase assaysas well as the need to specify the temperature at which theratio of activities for the enzyme are clearly established.
1Present Address: Cropping Systems Research Laboratory, USDA-ARS,Route 3, Box 215, Lubbock, TX 79401, U.S.A. (Received November 25, 1987; Accepted March 2, 1988) 相似文献
26.
A rapid fluorometric DNA assay for the measurement of cell density and proliferation in vitro 总被引:4,自引:0,他引:4
Timothy A. McCaffrey Lily A. Agarwal Babette B. Weksler 《In vitro cellular & developmental biology. Plant》1988,24(3):247-252
Summary Many research efforts require the accurate determination of cell density in vitro. However, physical cell counting is inaccurate,
time-intensive and requires removal of the cells from their growth environment, thereby introducing a host of potential artifacts.
The current studies document a very simple method of determining cell density in microtiter wells via DNA-enhanced fluorescence.
Fixed cells are stained with the A-T intercalating DNA stains DAPI or Hoechst 33342 and then fluorescence is quantified in
a plate fluorometer. Fluorescence is shown to be linearly related to cell density as determined by two physical counting methods.
The validity of the method is established in determining serum-stimulated growth of smooth muscle cells and in mitogen-induced
growth of endothelial cells. The fixed cells can be stored for prolonged periods, thus allowing time-course proliferation
assays without interassay variations. The fixed cells are also suitable for determinations of antigens of interest by ELISA.
This method is potentially valuable in many in vitro systems where the quantification of cell density and proliferation is
necessary.
This work supported in part by NIH Cardiovascular Training Grant HL07423 and a grant from the American Federation for Aging
Research to T. M. and HL35724 to B. W.
EDITOR’S STATEMENT The technique described in this paper represents an approach to quantifying cell density in adherent monolayers
of cultured cells in microtiter wells that is rapid and simple and does not require radioisotopes or removal of cells. 相似文献
27.
28.
Mauricio M. Bustos Fatma A. Kalkan Kathryn A. VandenBosch Timothy C. Hall 《Plant molecular biology》1991,16(3):381-395
An intron-less phaseolin gene [15] was used to express phaseolin polypeptides in transgenic tobacco plants. The corresponding amounts of phaseolin immunoreactive polypeptides and mRNA were similar to those found in plants transformed with a bean genomic DNA sequence that encodes an identical -phaseolin subunit. These results justified the use of the intron-less gene for engineering of the phaseolin protein by oligonucleotide-directed mutagenesis. Each and both of the two Asn residues that serve as glycan acceptors in wild-type phaseolin were modified to prevent N-linked glycosylation. Wild-type (wti–) and mutant phaseolin glycoforms (dgly
1, dgly
2 and dgly
1,2) were localized to the protein body matrix by immunogold microscopy. Although quantitative slot-blot hybridization analysis showed similar levels of phaseolin mRNA in transgenic seed derived from all constructs, seed from the dgly
1 and dgly
2 mutations contained only 41% and 73% of that expressed from the wild-type control; even less (23%) was present in seed of plants transformed with the phaseolin dgly
1,2 gene. Additionally, the profile of 25–29 kDa processed peptides was different for each of the glycoforms, indicating that processing of the full-length phaseolin polypeptides was modified. Thus, although targeting of phaseolin to the protein body was not eliminated by removal of the glycan side-chains, decreased accumulation and stability of the full-length phaseolin protein in transgenic tobacco seed were evident.Abbreviations bp
base pair(s)
- DAF
days after flowering
- GUS
-glucuronidase
- kb
kilobase
- kDa
kilodalton 相似文献
29.
Timothy J. Fahey 《Mycorrhiza》1992,1(2):83-89
Summary Mycorrhizae play an important role in regulating patterns of energy and nutrient flux in terrestrial ecosystems. To conceptualize this role I develop the theory behind a simple index of the efficiency of soil resource acquisition by plant root systems (E). The morphological, physiological and demographic characteristics of mycorrhizae that define E appear to vary with environment and with plant community composition. This theory is elaborated with examples drawn from forest ecology literature. Some inconsistencies among observations of fine root dynamics are particularly revealing: (1) belowground carbon allocation vs soil fertility; (2) causes of root mortality; (3) root longevity vs decomposition rates. A comprehensive theory of mycorrhizal and ecosystem dynamics must await resolution of these inconsistencies and better quantitative information on mycorrhizal features affecting E. 相似文献
30.
A homologue of the Escherichia coli DsbA protein involved in disulphide bond formation is required for enterotoxin biogenesis in Vibrio cholerae 总被引:22,自引:0,他引:22
A strain of Vibrio cholerae, which had been engineered to express high levels of the non-toxic B subunit (EtxB) of Escherichia coli heat-labile enterotoxin, was subjected to transposon (TnphoA) mutagenesis. Two chromosomal TnphoA insertion mutations of the strain were isolated that showed a severe defect in the amount of EtxB produced. The loci disrupted by TnphoA in the two mutant derivatives were cloned and sequenced, and this revealed that the transposon had inserted at different sites in the same gene. The open reading frame of the gene predicts a 200-amino-acid exported protein, with a Cys-X-X-Cys motif characteristic of thioredoxin, protein disulphide isomerase, and DsbA (a periplasmic protein required for disulphide bond formation in E. coli). The V. cholerae protein exhibited 40% identity with the DsbA protein of E. coli, including 90% identity in the region of the active-site motif. Introduction of a plasmid encoding E. coli DsbA into the V. cholerae TnphoA derivatives was found to restore enterotoxin formation, whilst expression of Etx or EtxB in a dsbA mutant of E. coli confirmed that DsbA is required for enterotoxin formation in E. coli. These results suggest that, since each EtxB subunit contains a single intramolecular disulphide bond, a transient intermolecular interaction with DsbA occurs during toxin subunit folding which catalyses formation of the disulphide in vivo. 相似文献