全文获取类型
收费全文 | 3023篇 |
免费 | 156篇 |
出版年
2021年 | 25篇 |
2019年 | 15篇 |
2018年 | 28篇 |
2017年 | 32篇 |
2016年 | 51篇 |
2015年 | 64篇 |
2014年 | 70篇 |
2013年 | 266篇 |
2012年 | 146篇 |
2011年 | 135篇 |
2010年 | 89篇 |
2009年 | 104篇 |
2008年 | 162篇 |
2007年 | 160篇 |
2006年 | 172篇 |
2005年 | 134篇 |
2004年 | 163篇 |
2003年 | 166篇 |
2002年 | 172篇 |
2001年 | 39篇 |
2000年 | 30篇 |
1999年 | 54篇 |
1998年 | 44篇 |
1997年 | 53篇 |
1996年 | 47篇 |
1995年 | 38篇 |
1994年 | 35篇 |
1993年 | 35篇 |
1992年 | 46篇 |
1991年 | 36篇 |
1990年 | 36篇 |
1989年 | 32篇 |
1988年 | 35篇 |
1987年 | 23篇 |
1986年 | 37篇 |
1985年 | 24篇 |
1984年 | 28篇 |
1983年 | 22篇 |
1982年 | 40篇 |
1981年 | 16篇 |
1980年 | 31篇 |
1979年 | 22篇 |
1978年 | 19篇 |
1977年 | 19篇 |
1976年 | 16篇 |
1975年 | 14篇 |
1974年 | 14篇 |
1973年 | 15篇 |
1970年 | 19篇 |
1969年 | 17篇 |
排序方式: 共有3179条查询结果,搜索用时 15 毫秒
11.
Summary In a continuous culture of Bacillus caldolyticus strain SP, which requires maltose as an inducer for production of -amylase in batch culture, a predominant mutant strain M1 which produced high amounts of -amylase in the absence of maltose in batch culture, developed. The change of cell population from strain SP to strain M1 in maltose-casitone medium was linear with time in the transient state after the change from batch to continuous culture at a dilution rate of 0.17 h-1, and was completed in about 11 generations of bacterial growth. The dilution rate effect of continuous culture on -amylase activity was almost the same with both strains SP and M1. The maximum -amylase activity of 380 units/ml was observed at an intermediate dilution rate that was 11.5 times higher than -amylase activity at the end of a batch culture using the same medium. It was deduced that the enhancement of -amylase production in continuous culture was attributed partly to the predominant growth of a mutant strain with higher -amylase productivity. 相似文献
12.
Summary In an attempt to reveal the role of antigen-laden marginal metallophil (MM) and other macrophages in the intrasplenic immune response of a specific B-cell lineage to a thymus-independent type-2 antigen (Ficoll conjugated with fluorescein isothiocyanate), simultaneous immuno-histological observations of the involved cells were performed in the rat. By newly established methods of double or triple immunostainings, time-kinetics of the following parameters were studied and compared: (1) the antigen, (2) the specific antibody-forming cells (AFC) directed to the fluorescein-isothiocyanate determinant, (3) proliferating cells labeled with 5-bromo-2-deoxyuridine (BrdU), and (4) macrophage subpopulations recognized by monoclonal antibodies (ED2 and ED3). The antigen localized stably not only in the marginal-zone macrophages but also in the MM except around the follicular area. The increase of BrdU-positive cells was observed from day 2 up to day 4 after antigen injection mostly in the periphery of the periarterial lymphoid sheath (outer PALS), which indicated antigen-induced proliferation. As a novel finding, the majority of AFC, both BrdU-positive and -negative, were either closely associated with the antigen-laden MM, or forming cell clusters with ED2-positive macrophages in the outer PALS. In contrast, there were very few AFC in juxtaposition to antigen-free MM in the follicular area or the antigen-laden marginal zone macrophages. The results led to the proposal of a hypothesis that the antigen-laden MM together with ED2-positive macrophages constitute an immunoproliferative microenvironment for the plasmacellular reaction by accumulating the antigen-specific B-cell lineage and promoting these cells to differentiate into the AFC and to proliferate in the outer PALS.Abbreviations
AFC
specific antibody-forming cells
- BrdU
5-bromo-2-deoxyuridine
- Fic-F
FITC-conjugated Ficoll
- FITC
fluorescein isothiocyanate
- HRP
horseradish peroxidase
- MM
marginal metallophils
- MZ
marginal zone
- PALS
periarterial lymphoid sheath
- PBS
phosphate-buffered saline
- TI2
thymus-independent type-2 相似文献
13.
Scale Arrangement Pattern in a Lepidopteran Wing. 1. Periodic Cellular Pattern in the Pupal Wing of Pieris rapae 总被引:1,自引:1,他引:0
In most species of lepidopteran insects, anteroposterior rows formed by scales are arranged at regular intervals in the adult wing; within each row two kinds of scales are alternately arranged. To investigate the cellular basis for the scale arrangement pattern, we examined cell arrangement in the epidermal monolayer of the pupal wing of a small white cabbage butterfly, Pieris rapae , by scanning electron microscopy and light microscopy.
The arrangement of scale precursor cells, closely resembling that of scales in the adult wing, was observed in the wing epidermis of the early pupa. Scale precursor cells are proximodistally elongated and form anteroposterior rows. Within a row two kinds of scale precursor cells are nearly alternately arranged, which is not so precise as the alternation of scales in the adult wing. Individual rows of scale precursor cells are separated by rows of single or double undifferentiated general epidermal cells. Occasionally, arrangement abnormalities occur both in the adult and the pupal wing. The cellular basis for the regular spacing of scale rows is discussed. 相似文献
The arrangement of scale precursor cells, closely resembling that of scales in the adult wing, was observed in the wing epidermis of the early pupa. Scale precursor cells are proximodistally elongated and form anteroposterior rows. Within a row two kinds of scale precursor cells are nearly alternately arranged, which is not so precise as the alternation of scales in the adult wing. Individual rows of scale precursor cells are separated by rows of single or double undifferentiated general epidermal cells. Occasionally, arrangement abnormalities occur both in the adult and the pupal wing. The cellular basis for the regular spacing of scale rows is discussed. 相似文献
14.
Isolation and Characterization of an Enterobacter cloacae Strain That Reduces Hexavalent Chromium under Anaerobic Conditions 总被引:12,自引:6,他引:6
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Pi-Chao Wang Tsukasa Mori Kohya Komori Masanori Sasatsu Kiyoshi Toda Hisao Ohtake 《Applied microbiology》1989,55(7):1665-1669
An Enterobacter cloacae strain (HO1) capable of reducing hexavalent chromium (chromate) was isolated from activated sludge. This bacterium was resistant to chromate under both aerobic and anaerobic conditions. Only the anaerobic culture of the E. cloacae isolate showed chromate reduction. In the anaerobic culture, yellow turned white with chromate and the turbidity increased as the reduction proceeded, suggesting that insoluble chromium hydroxide was formed. E. cloacae is likely to utilize toxic chromate as an electron acceptor anaerobically because (i) the anaerobic growth of E. cloacae HO1 accompanied the decrease of toxic chromate in culture medium, (ii) the chromate-reducing activity was rapidly inhibited by oxygen, and (iii) the reduction occurred more rapidly in glycerol- or acetate-grown cells than in glucose-grown cells. The chromate reduction in E. cloacae HO1 was observed at pH 6.0 to 8.5 (optimum pH, 7.0) and at 10 to 40°C (optimum, 30°C). 相似文献
15.
Mutational analysis of structure--activity relationships in human tumor necrosis factor-alpha 总被引:10,自引:0,他引:10
J Yamagishi H Kawashima N Matsuo M Ohue M Yamayoshi T Fukui H Kotani R Furuta K Nakano M Yamada 《Protein engineering》1990,3(8):713-719
To determine the region of human tumor necrosis factor-alpha (TNF-alpha), essential for cytotoxic activity against mouse L-M cells, single amino-acid-substituted TNF-alpha mutant proteins (muteins) were produced in Escherichia coli by protein engineering techniques. An expression plasmid for TNF-alpha was mutagenized by passage through an E. coli mutD5 mutator strain and by oligonucleotide-directed mutagenesis. Approximately 100 single amino-acid-substituted TNF-alpha muteins were produced and assayed for cytotoxic activity. The cytotoxic activities of purified TNF-alpha muteins, e.g. TNF-31T, -32Y, -82D, -85H, -115L, -141Y, -144K and -146E, were less than 1% of that of parent TNF-alpha. These results indicate that the integrity of at least four distinct regions of the TNF-alpha molecule is required for full biological activity. These regions are designated as follows: region I, from position 30 to 32; region II, from position 82 to 89; region III, from position 115 to 117; region IV, from position 141 to 146. In addition, TNF-141Y could not completely compete with parent TNF-alpha for binding to the receptor. This demonstrates that region IV, and at least aspartic acid at position 141, must be involved in the TNF receptor binding site. 相似文献
16.
Yuzuru Mikami Sen Feng Yu Katsukiyo Yazawa Kazutaka Fukushima Akio Maeda Jun Uno Kiyoshi Terao Naoki Saito Akinori Kubo Ken -ichiro Suzuki 《Mycopathologia》1990,112(2):113-118
During our studies on toxic substances from clinically isolated Nocarida, a new isolate identified as Nocardia otitidiscaviarum from cutaneous nocardiosis was found to produce a toxic substance called HS-6 that had strong in vitro as well as in vivo toxicity. The mouse intraperitoneal LD50 value was 1.25 mg/kg and the ED50 value for L1210 cultured cells was 0.3 ng/ml. The structure of HS-6 was determined and found to belong to the 16-membered macrocyclic group with a molecular formula of C43H68O12. HS-6 also showed activity against pathogenic fungi such as Cryptococcus neoformans. 相似文献
17.
Cloning, nucleotide sequence, and expression of the HincII restriction-modification system. 总被引:10,自引:9,他引:1
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Two genes, coding for the HincII from Haemophilus influenzae Rc restriction-modification system, were cloned and expressed in Escherichia coli RR1. Their DNA sequences were determined. The HincII methylase (M.HincII) gene was 1,506 base pairs (bp) long, corresponding to a protein of 502 amino acid residues (Mr = 55,330). The HincII endonuclease (R.HincII) gene was 774 bp long, corresponding to a protein of 258 amino acid residues (Mr = 28,490). The amino acid residues predicted from the R.HincII and the N-terminal amino acid sequence of the enzyme found by analysis were identical. These methylase and endonuclease genes overlapped by 1 bp on the H. influenzae Rc chromosomal DNA. The clone, named E. coli RR1-Hinc, overproduced R.HincII. The R.HincII activity of this clone was 1,000-fold that from H. influenzae Rc. The amino acid sequence of M.HincII was compared with the sequences of four other adenine-specific type II methylases. Important homology was found between tne M.HincII and these other methylases. 相似文献
18.
19.
Nobutaka Fujii Akira Otaka Susumu Funakoshi Toshihiro Watanabe Hiromitsu Arai Kiyoshi Bessho Haruaki Yajima 《Journal of Protein Chemistry》1988,7(2):151-156
Treatment of a mixture of Cys(R)(O) and Cys(R) with an acid was found to generate cystine in fairly good yields, when suitable R, R, and an acid were selected. An unsymmetrical cystine peptide was prepared by treatment of a mixture of Z(OMe)-Cys(R) (0)-Ala-NH2 (R=Acm or MBzl) and Z(OMe)-Cys(MBzl)-Gly-OBzl with TFA or 1 M TFMSA/TFA.3 Oxytocin was obtained in an excellent yield by TFA treatment of the protected peptide containing Cys(Acm)(0) and Cys(MBzl). Thus, formation of the disulfide bond was found feasible at the position of Cys(R) (0).The following abbreviations are used Boc
t-butyloxycarbonyl
- Z(OMe)
p-methoxybenzyloxycarbonyl
- MBzl
p-methoxybenzyl
- Acm
acetamidomethyl
- Bzl
benzyl
- Ad
l-adamantyl
- tBu
t-butyl
- TFA
trifluoroacetic acid
- TFMSA
trifluoromethanesulfonic acid
- TMSOTf
trimethylsilyl trifluoromethane sulfonate 相似文献
20.
Dr. Emiko Matsumoto Kazushige Hirosawa Kiyoshi Takagawa Yoshiki Hotta 《Cell and tissue research》1988,252(2):293-300
Summary A Drosophila visual mutant rdgA has photoreceptive cells which degenerate gradually after eclosion. Fine structure of the retinular cells of rdgA
KS60
and rdgA
K014
was studied during early stages of degeneration to determine the initial morphological defects. The retinular cells of these two alleles showed the following structural abnormality within 1 day after eclosion: (1) rhabdomeres were small and irregular in shape; (2) cisternae of the rough endoplasmic reticulum were more numerous than those in normal retinular cells; (3) submicrovillar cisternae were absent; and (4) lysosomes were fewer than normal. Three-dimensional reconstruction of serial sections of the ommatidia showed that the degeneration of mutant rhabdomeres proceeds more rapidly in regions remote from the nuclei. These results suggest that the process of turnover of rhabdomeric microvilli is abnormal in rdgA. We also confirmed an increase of lysosomes and destruction of cellular organelles, as reported by previous investigators at more advanced stages of degeneration. 相似文献