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31.
The 'two-step' model proposed by Jensen and his collaborators for explaining estrogen action conceptualized hormone-free estrogen receptors (ER) to be cytoplasmic, and hormone-filled, transformed ER to be nuclear. Applying monoclonal antibodies which recognized epitopes in ER and formaldehyde-fixed tissues, King et al demonstrated exclusively nuclear staining in target tissues utilizing immunoperoxidase technique. Recently these antibodies have become commercially available enabling other investigators to conduct studies. In this report, using these monoclonal antibodies we have demonstrated that a change in the concentration of formaldehyde alters the staining pattern yielding cytoplasmic instead of nuclear staining in calf uterus, MCF-7 cells, and ER(+) human breast cancer. In addition, neutralization of the antibody activity was not achieved with freshly prepared ER(+) cytosols. Formaldehyde-treated cytosols were essential. These results ought to caution investigators in determining in vivo location of antigens based on the staining pattern obtained in fixed tissues. Furthermore, this effect of formaldehyde on estrogen receptors may be applicable to other steroid hormone receptors. 相似文献
32.
33.
Kiyoshi Takahashi Katsuya Miyatani Hiroyuki Yanai Ho Jong Jeon Kotaro Fujiwara Tadashi Yoshino Kazuhiko Hayashi Tadaatsu Akagi Ken Tsutsui Koichi Mizobuchi 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,62(1):105-113
Monocytic leukemia (MoL) cells were obtained from the peripheral blood of a patient in whom the leukemic cells infiltrating
various lymphoreticular organs exhibited features intermediate between interdigitating reticulum cells (IDC) and ordinary
phagocytic macrophages, whereas the leukemic cells in the peripheral blood were essentially monocytic and lacked such features.
Peripheral blood CD4+ T-cells were established as an interleukin-2-dependent T-cell line. When the MoL cells were exposed
for a few days to conditioned medium from the T-cell line, they extended several dendritic cytoplasmic projections and became
intensely positive for HLA-DR antigen, cytoplasmic S-100β protein, and CD1 antigen. Functionally, the conditioned medium significantly
down-regulated Fc-mediated and Fc-independent phagocytic activities, and the levels of lysosomal enzymes such as lysozyme
and nonspecific esterase in the MoL cells. Moreover, the conditioned medium significantly up-regulated the accessory cell
function of the MoL cells as measured by the primary allogenic mixed leukocyte reaction (MLR). Furthermore, the conditioned
medium significantly down-regulated the expression of CD14 antigen.
Biochemical analysis indicated that the factor responsible for these changes is a protein which is distinct from known human
cytokines and whose molecular weight is approximately 31 kDa. These findings suggest that IDC are closely related the monocytic
lineage and that helper T-cells play an important role in constructing the microenvironment of T-lymphoid tissues which is
necessary for the differentiation and maturation of IDC. 相似文献
34.
K Tamura 《Molecular biology and evolution》1992,9(5):814-825
The nucleotide sequences of a segment of mitochondrial DNA (mtDNA) have been determined for nine species or subspecies of the subgenus Drosophila of the genus Drosophila. This segment contains two complete protein-coding genes (i.e., NADH dehydrogenase subunit 1 and cytochrome b) and a transfer RNA gene (tRNA(ser)). The G+C content at third-codon positions for the two protein-coding genes was 1.5 times higher than that in the D. melanogaster species group, which belongs to the subgenus Sophophora. However, there was a substantial difference between the nucleotide frequencies of G and C. The number of nucleotide substitutions per silent site was more than three times higher than that for nuclear DNA, although it was only 60% of that for mammalian mtDNA. Both parametric and nonparametric analyses revealed a strong transition-transversion bias in nucleotide substitution, as was observed in mammalian mtDNA. Moreover, the rate of substitution of A and T for G and C is higher than that for the opposite direction. This bias seems to be responsible for the extremely A+T-rich base composition of Drosophila mtDNA. It is also noted that the rate of transitional change between A and G is higher than that between T and C. 相似文献
35.
Cloning of the alpha-amylase cDNA of Aspergillus shirousamii and its expression in Saccharomyces cerevisiae. 总被引:2,自引:0,他引:2
I Shibuya G Tamura T Ishikawa S Hara 《Bioscience, biotechnology, and biochemistry》1992,56(2):174-179
alpha-Amylase cDNA was cloned and sequenced from Aspergillus shirousamii RIB2504. The putative protein deduced from the cDNA open reading frame (ORF) consisted of 499 amino acids with a molecular weight of 55,000. The amino acid sequence was identical to that of the ORF of the Taka-amylase A gene of Aspergillus oryzae, while the nucleotide sequence was different at two and six positions in the cDNA ORF and 3' non-coding regions, respectively, so far determined. The alpha-amylase cDNA was expressed in Saccharomyces cerevisiae under the control of the yeast ADH1 promoter using a YEp-type plasmid, pYcDE1. The cDNA of glucoamylase, which was previously cloned from the same organism, was also expressed under the same conditions. Consequently, active alpha-amylase and glucoamylase were efficiently secreted into the culture medium. The amino acid sequence of the N-terminal regions of these enzymes purified from the yeast culture medium confirmed that the signal sequences of these enzymes were cleaved off at the same positions as those of the native enzymes of A. shirousamii. 相似文献
36.
K Tsuchiya S Tada K Gomi K Kitamoto C Kumagai G Tamura 《Bioscience, biotechnology, and biochemistry》1992,56(11):1849-1853
The Taka-amylase A gene (amyB) of Aspergillus oryzae is induced by starch or maltose. The molecular mechanism of the induction was investigated using a fusion of the amyB promoter and the Escherichia coli uidA gene encoding beta-glucuronidase (GUS). To identify the region responsible for high-level expression and regulation within the amyB promoter, a series of deletion promoters was constructed and introduced into the A. oryzae met locus by homologous recombination. Deletion of the region between -377 to -290 (the number indicates the distance in base pairs from the translation initiation point (+1) to the deletion end point) significantly reduced of the GUS activity, but slight reduction of the GUS activity was observed in deletions up to -377. Northern blot analysis showed that reduction of the GUS activity depended upon the expression level of the GUS gene. The region between -377 to -290 is suggested to include the sequence required directly for high-level expression and regulation of the amyB gene. 相似文献
37.
Setsuzo Tada Katsuya Gomi Katsuhiko Kitamoto Kojiro Takahashi Gakuzo Tamura Shodo Hara 《Molecular & general genetics : MGG》1991,229(2):301-306
Summary Northern blot analysis of glucose-grown and starch-grown mycelia of Aspergillus oryzae R11340 was conducted using the cloned Taka-amylase A (TAA) gene as a probe. The amount of mRNA homologous to the TAA gene was increased when this fungus was grown with starch as a sole carbon source. In order to analyze the induction mechanism, we inserted the Escherichia coli uidA gene encoding -glucuronidase (GUS) downstream of the TAA promoter and introduced the resultant fusion gene into the A. oryzae genome. Production of a functional GUS protein was induced by starch, but not by glucose. When the effects of various sugars on expression of the fusion gene were examined, the results suggested that the expression of the fusion gene was under control of the TAA gene promoter. 相似文献
38.
The second-order rate constant (k4) for the oxidation of monosubstituted phenols and anilines by lactoperoxidase compound II was examined by Chance's method [B. Chance, Arch. Biochem. Biophys.
71 (1957), 130–136]. When the electronic states of these substrates were calculated by an ab initio molecular orbital method, it was found that the log k4 value correlates well with the highest occupied molecular orbital (HOMO) energy level but not with the net charge or frontier electron density. These results are essentially similar to those reported previously in the case of horseradish peroxidase [J. Sakurada, R. Sekiguchi, K. Sato, and T. Hosoya, Biochemistry
29 (1990), 4093–4098], showing some dissimilar features which are considered to reflect the structural difference between the two enzymes.Abbreviations HOMO
highest occupied molecular orbital
- HRP
horseradish peroxidase
- LPO
lactoperoxidase (EC 1.11.1.7)
- LUMO
lowest unoccupied molecular orbital 相似文献
39.
A repeating unit of the DYZ1 family on the human Y chromosome consists of segments with partial male-specificity 总被引:1,自引:0,他引:1
Y Nakagome S Nagafuchi S Seki Y Nakahori T Tamura M Yamada M Iwaya 《Cytogenetics and cell genetics》1991,56(2):74-77
An average-sized human Y chromosome contains about 3,000 copies of the repeating DNA family DYZ1. A major repeating unit of the family, pHY10, has been cloned and an entire 3,564-bp sequence has already been determined by Nakahori et al. (1986). In the present study, pHY10 was divided into six consecutive segments, A to F, which were independently amplified by the PCR technique to see if they were strictly male-specific. pHY10 appears to consist of segments of various male-specificity. The B segment was apparently male-specific; however, the use of additional techniques (Southern-blot analysis or second PCR amplification in combination with the standard PCR) revealed homologous sequences in some females. None of the six segments of pHY10 may be male-specific in a strict sense. Different segments appear to be conserved during evolution to different extents. The 323-bp E segment appears to be the least conserved and to be responsible for the generation of most variations within the DYZ1 family. 相似文献
40.
Striking homology of the ''variable'' N-terminal as well as the ''conserved core'' domains of the mouse and human TATA-factors (TFIID). 总被引:18,自引:9,他引:9 下载免费PDF全文
T Tamura K Sumita I Fujino A Aoyama M Horikoshi A Hoffmann R G Roeder M Muramatsu K Mikoshiba 《Nucleic acids research》1991,19(14):3861-3865