全文获取类型
收费全文 | 19703篇 |
免费 | 1327篇 |
国内免费 | 7篇 |
专业分类
21037篇 |
出版年
2024年 | 18篇 |
2023年 | 132篇 |
2022年 | 120篇 |
2021年 | 221篇 |
2020年 | 267篇 |
2019年 | 244篇 |
2018年 | 603篇 |
2017年 | 515篇 |
2016年 | 791篇 |
2015年 | 1169篇 |
2014年 | 1095篇 |
2013年 | 1505篇 |
2012年 | 1795篇 |
2011年 | 1686篇 |
2010年 | 1004篇 |
2009年 | 795篇 |
2008年 | 1256篇 |
2007年 | 1198篇 |
2006年 | 1157篇 |
2005年 | 999篇 |
2004年 | 1008篇 |
2003年 | 890篇 |
2002年 | 835篇 |
2001年 | 137篇 |
2000年 | 99篇 |
1999年 | 136篇 |
1998年 | 126篇 |
1997年 | 81篇 |
1996年 | 97篇 |
1995年 | 94篇 |
1994年 | 84篇 |
1993年 | 85篇 |
1992年 | 74篇 |
1991年 | 56篇 |
1990年 | 48篇 |
1989年 | 43篇 |
1988年 | 39篇 |
1987年 | 30篇 |
1986年 | 24篇 |
1985年 | 43篇 |
1984年 | 48篇 |
1983年 | 27篇 |
1982年 | 39篇 |
1981年 | 21篇 |
1980年 | 19篇 |
1979年 | 21篇 |
1978年 | 17篇 |
1977年 | 12篇 |
1976年 | 28篇 |
1975年 | 22篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
11.
Marco L. F. Giuseppin Hendrikus M. J. van Eijk Marja Hellendoorn José W. van Almkerk 《Applied microbiology and biotechnology》1987,27(1):31-36
Summary The changes in cell wall strength of Hansenula polymorpha have been investigated in continuous cultures with respect to the recovery of methanol oxidase (MOX). Cultures grown on several substrate mixtures that enable induction of MOX have been compared with cultures grown on methanol as the sole inducer. The effects of dilution rate (D) on lysis properties have been studied. The cell wall strength was consistently influenced by growth media and D. Media containing glycerol/methanol showed the slowest lysis kinetics, with a large fraction of non-degradable cell wall material. In continuous cultures grown on a mixture of glucose and methanol both the resistance to zymolyase and the mean cell wall thickness increased at D<0.1 h–1. The yield of MOX by zymolyase lysis is reproducible and up to 100% higher than that of the standard ultrasonic treatment. The lysis kinetics indicated that zymolyase punctures the cell wall; since the release rate of MOX is lower than that of protein, the cell contents will leak through. At D-values>0.2 h–1, both protein and MOX release rates increase, reflecting a change in lysis mechanism due to the increased fraction of thin daughter cells. Kinetic analysis of zymolyase lysis using both physical and enzymatic methods provides information for achieving optimal recovery of MOX.Abbreviations and symbols
C
MOX
MOX activity [MOX units·g X–1]
-
D
dilution rate [h–1]
- MOX
methanol oxidase
- kc
decay rate constant of A 610 nm [min–1]
- kd
decay constant of MOX activity [min–1]
- kdis
dissociation rate constant [min–1]
- kMOX
release rate constant of MOX activity [min–1]
- kp
release rate constant of protein [min–1]
- R
recovery efficiency of enzyme [-]
-
St
stability of enzyme [-] 相似文献
12.
We have studied the chromatin structure ofPenicillium chrysogenum. This fungus presents the typical nucleosomal repeat and the core DNA size characteristic of all the eukaryotes. The repeat length (about 180 base pairs) is in the range of those obtained for most fungi (160–180 base pairs) and shorter than in higher eukaryotes. Knowledge aboutP. chrysogenum chromatin structure opens the way to the study of the mechanisms of genetic regulation in this filamentous fungus. 相似文献
13.
The in situ localization of Ca2+ in stage I sporangiophores of the fungus Phycomyces blakesleeanus was achieved with the potassium pyroantimonate technique. Precipitates of calcium-antimonate were present in mitochondria, vacuoles, endoplasmic reticulum and adjacent cytoplasm, Golgi-like bodies, and nuclei but not cell walls. Material treated with the calcium chelator EGTA lacked these precipitates. The preferential localization of Ca2+ in mitochondria, endoplasmic reticulum and vacuoles suggests that these organelles modulate the level of this cation in sporangiophores of P. blakesleeanus.Abbreviations EGTA
ethyleneglycol-bis-(-aminoethyl ether) N,N, tetraacetic acid 相似文献
14.
On the solution of mathematical models of herd immunity in human helminth infections 总被引:1,自引:0,他引:1
Marco V. José 《Journal of mathematical biology》1989,27(6):707-715
The general solution of the mathematical model of herd immunity to human helminth infections recently proposed by Anderson and May [3] is obtained. The numerical solution of a more accurate biological model is indistinguishable from the corresponding exact solution of a more tractable mathematical model. Computer simulations of some particular cases of this model support the notion that both ecological and immunological factors determine the observed convex patterns of age-prevalence and age-intensity curves of human helminth infections.This work was made thanks to the advise and support of Dr. Robert M. May while the author was Postdoctoral Fellow at Princeton University 相似文献
15.
Josée Harel Linda Duplessis Jeffrey S. Kahn Michael S. DuBow 《Archives of microbiology》1990,154(1):67-72
The 37,000 bp double-stranded DNA genome of bacteriophage Mu behaves as a plaque-forming transposable element of Escherichia coli. We have defined the cis-acting DNA sequences required in vivo for transposition and packaging of the viral genome by monitoring the transposition and maturation of Mu DNA-containing pSC101 and pBR322 plasmids with an induced helper Mu prophage to provide the trans-acting functions. We found that nucleotides 1 to 54 of the Mu left end define an essential domain for transposition, and that sequences between nucleotides 126 and 203, and between 203 and 1,699, define two auxiliary domains that stimulate transposition in vivo. At the right extremity, the essential sequences for transposition require not more than the first 62 base pairs (bp), although the presence of sequences between 63 and 117 bp from the right end increases the transposition frequency about 15-fold in our system. Finally, we have delineated the pac recognition site for DNA maturation to nucleotides 32 to 54 of the Mu left end which reside inside of the first transposase binding site (L1) located between nucleotides 1–30. Thus, the transposase binding site and packaging domains of bacteriophage Mu DNA can be separated into two well-defined regions which do not appear to overlap.Abbreviations
attL
attachment site left
-
attR
attachment site right
- bp
base pairs
- Kb
kilobase pair
- nt
nucleotide
- Pu
Purine
- Py
pyrimidine
- Tn
transposable element
State University of New York, Downstate Medical Center, Brooklyn, NY 11204 USA 相似文献
16.
Raúl Castaño Angel Ezquerra Nieves Doménech José A. López de Castro 《Immunogenetics》1988,27(5):345-355
The HLA-A2 antigen expressed by donor OZB can be distinguished from the main HLA-A2.1 subtype by isoelectric focusing - it is one charge unit more acidic — and by some alloreactive T-cell clones but not by cytolytic T lymphocyte lines. The structure of variant OZB has been examined by comparative peptide mapping with A2.1 and radiochemical sequence analysis. The two molecules were found to differ in a single tryptic peptide from the 0 region, spanning residues 220–243. The amino acid sequence of this peptide from variant OZB revealed that there was only one amino acid change of Glu instead of Ala at position 236, a hitherto invariant residue in class I HLA antigens. All previously characterized HLA or H-2 natural variants have structural changes restricted to the 1 and/or 2 domains. Thus, variant OZB is unique in that (1) it has one amino acid change in 3 and (2) it has no changes in l and 2. The only detected substitution of this variant may be accounted for by a single base change at the DNA level, suggesting that it might have resulted from a point mutation in the A2.1 gene. The structural features of variant OZB open a novel way to examine the influence of polymorphism in 3 on cytolytic T-cell recognition of naturally occurring class I antigens.Abbreviations CTL
cytolytic T lymphocytes
- HPLC
high performance liquid chromatography
- IEF
isoelectric focusing
- MHC
major histocompatibility complex 相似文献
17.
Ronald E. Koes Cornelis E. Spelt Jos N. M. Mol Anton G. M. Gerats 《Plant molecular biology》1988,10(4):375-385
Chalcone synthase (CHS) genes in Petunia hybrida comprise a multigene family containing at least 7 complete members in the strain Violet 30 (V30). Based on a high sequence homology in both coding and non-coding sequence, a number of CHS genes can be placed into two subfamilies. By restriction fragment length polymorphism (RFLP) analysis it was shown that both chromosomes II and V carry one of these subfamilies, in addition to the other CHS genes identified so far. Members of a subfamily were found to be closely linked genetically. Analysis of the Petunia species that contributed to the hybrid nature of P. hybrida (P. axillaris, P. parodii, P. inflata and P. violacea) shows that none of the CHS gene clusters is specific for either one of the parents and therefore did not arise as a consequence of the hybridization. The number of CHS genes within a subfamily varies considerably among these Petunia species. From this we infer that the CHS subfamilies arose from very recent gene duplications. 相似文献
18.
José M. Facelli 《Plant Ecology》1988,78(1-2):21-25
This paper reports on changes induced by the introduction of cattle in a grassland that had remained ungrazed for 9 yr, in comparison with two adjacent grasslands: one that remained enclosed and one that has been continuously subject to grazing. Basal cover was measured on 25 interception lines, each 1 m long, three times during one year. The variables studied were: total cover, cover of grasses and dicots, cover of creeping grasses, floristic composition, and dissimilarity among sites. At the first sampling, 2 yr after cattle re-introduction, the newly grazed site was more similar to the ungrazed than to the grazed site. The newly grazed site had very low cover of dicots; the species of dicots present were different from those found in the continuously grazed area. Creeping grasses had higher cover in the newly grazed site than in the other sites, and continued to increase. At the last sampling, one year later, the newly grazed site had become more similar to the contiuously grazed site. Only after 5 yr of cattle grazing the exotic dicots that were dominant in the continuously grazed site, were recorded in the re-opened site. The absence of propagules of these species or the absence of safe sites may account for this delayed invasion. 相似文献
19.
Glutathione reductase fromSaccharomyces cerevisiae undergoes redox interconversionin situ andin vivo
José Peinado Javier Florindo J. López-Barea 《Molecular and cellular biochemistry》1992,110(2):135-143
Redox interconversion of glutathione reductase was studiedin situ withS. cerevisiae. The enzyme was more sensitive to redox inactivation in 24 hour-starved cells than in freshly-grown ones. While 5 μM NADPH
or 100 μM NADH caused 50% inactivation in normal cells in 30 min, 0.75 μM NADPH or 50 μM NADH promoted a similar effect in
starved cells. GSSG reactivated the enzyme previously inactivated by NADPH, ascertaining that the enzyme was subjected to
redox interconversion. Low EDTA concentrations fully protected the enzyme from NADPH inactivation, thus confirming the participation
of metals in such a process. Extensive inactivation was obtained in permeabilized cells incubated with glucose-6-phosphate
or 6-phosphogluconate, in agreement with the very high specific activities of the corresponding dehydrogenases. Some inactivation
was also observed with malate, L-lactate, gluconate or isocitrate in the presence of low NADP+ concentrations.
The inactivation of yeast glutathione reductase has also been studiedin vivo. The activity decreased to 75% after 2 hours of growth with glucono-δ-lactone as carbon source, while NADPH rose to 144%
and NADP+ fell to 86% of their initial values. Greater changes were observed in the presence of 1.5 μM rotenone: enzymatic activity
descended to 23% of the control value, while the NADH/NAD+ and NADPH/NADP+ ratios rose to 171% and 262% of their initial values, respectively. Such results indicate that the lowered redox potential
of the pyridine nucleotide pool existing when glucono-δ-lactone is oxidized promotesin vivo inactivation of glutathione reductase. 相似文献
20.
Cecilia Zazueta José A. Holguín Jorge Ramírez 《Journal of bioenergetics and biomembranes》1991,23(6):889-902
We describe a calcium transport that is sensitive to ruthenium red in liposomes reconstituted with mitochondrial extracts. This system is able to build an internally negative membrane potential, which allows the electrogenic influx of Ca2+ and Sr2+. Proteins with molecular weights higher than 35 kDa were incorporated to the vesicles, and enhanced the accumulation of the cation in an energy-dependent fashion. 相似文献