全文获取类型
收费全文 | 777977篇 |
免费 | 94767篇 |
国内免费 | 403篇 |
出版年
2016年 | 8510篇 |
2015年 | 12963篇 |
2014年 | 14923篇 |
2013年 | 20776篇 |
2012年 | 23728篇 |
2011年 | 23671篇 |
2010年 | 16033篇 |
2009年 | 15242篇 |
2008年 | 21714篇 |
2007年 | 22310篇 |
2006年 | 20875篇 |
2005年 | 20136篇 |
2004年 | 19955篇 |
2003年 | 19313篇 |
2002年 | 18739篇 |
2001年 | 34296篇 |
2000年 | 34734篇 |
1999年 | 27820篇 |
1998年 | 10397篇 |
1997年 | 11037篇 |
1996年 | 10563篇 |
1995年 | 10316篇 |
1994年 | 10169篇 |
1993年 | 9996篇 |
1992年 | 23573篇 |
1991年 | 22839篇 |
1990年 | 22466篇 |
1989年 | 21797篇 |
1988年 | 20074篇 |
1987年 | 19730篇 |
1986年 | 18099篇 |
1985年 | 18180篇 |
1984年 | 15108篇 |
1983年 | 13188篇 |
1982年 | 10548篇 |
1981年 | 9384篇 |
1980年 | 8944篇 |
1979年 | 14539篇 |
1978年 | 11681篇 |
1977年 | 10558篇 |
1976年 | 9953篇 |
1975年 | 10796篇 |
1974年 | 11618篇 |
1973年 | 11407篇 |
1972年 | 10212篇 |
1971年 | 9475篇 |
1970年 | 8105篇 |
1969年 | 7660篇 |
1968年 | 6839篇 |
1967年 | 6170篇 |
排序方式: 共有10000条查询结果,搜索用时 31 毫秒
21.
22.
A partial swine cDNA which encodes the functional domain of PIT-1 was isolated by the polymerse chain reaction (PCR). The swine PIT-1 cDNA clone is 95% identical at the protein level to the rat Pit-1 gene. Thus, Pit-l's known function in control of rat growth hormone and prolactin expression is likely to be conserved in swine. This swine cDNA clone was used to investigate genetic variability at PIT-1 in several American and Chinese breeds. Polymorphic BamIII fragments were found in pure-bred Meishan animals (n= 13), but only monomorphic fragments in five American breeds (n= 36). 相似文献
23.
24.
Enzymatically active human testis angiotensin-converting enzyme (ACE) was expressed in Chinese hamster ovary (CHO) cells stably transfected with each of three vectors: p omega-ACE contains a full-length testis ACE cDNA under the control of a retroviral promoter; and pLEN-ACEVII and pLEN-ACE6/5, in which full-length and membrane anchor-minus testis ACE cDNAs, respectively, are under the control of the human metallothionein IIA promoter and SV40 enhancer. In every case, active recombinant human testis ACE (hTACE) was secreted in a soluble form into the culture media, up to 2.4 mg/liter in the media of metal-induced, high-producing clones transfected with one of the pLEN vectors. In addition, membrane-bound recombinant enzyme was recovered from detergent extracts of cell pellets of CHO cells transfected with either p omega-ACE or pLEN-ACE-VII. Recombinant converting enzyme was purified to homogeneity by single-step affinity chromatography of conditioned media and detergent-extracted cell pellets in 85 and 70% overall yield, respectively. Purified hTACE from all sources comigrated with the native testis isozyme on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with M(r) approximately 100 kDa. The native and recombinant proteins cross-reacted equally with anti-human kidney ACE antiserum on Western blotting. The catalytic activity of recombinant angiotensin-converting enzyme, in terms of angiotensin I and 2-furanacryloyl-Phe-Gly-Gly hydrolysis, chloride activation, and lisinopril inhibition, was essentially identical to that of the native enzyme. The facile recovery in high yield of fully active hTACE from the media of stably transfected CHO cells provides a suitable system for investigating structure-function relationships in this enzyme. 相似文献
25.
Jonathan H. Clarke Judith I. Laurie Harry J. Gilbert Geoffrey P. Hazlewood 《FEMS microbiology letters》1991,83(3):305-309
Cellulomonas fimi genomic DNA encoding xylanase activity has been cloned and expressed in Escherichia coli. As judged by DNA hybridization and restriction analysis, twelve xylanase-positive clones carried a minimum of four different xylanase (xyn) genes. The encoded enzymes were devoid of cellulase activity but three of the four bound to Avicel. 相似文献
26.
D R Hickey K Jayaraman C T Goodhue J Shah S A Fingar J M Clements Y Hosokawa S Tsunasawa F Sherman 《Gene》1991,105(1):73-81
Genes encoding tuna, pigeon, and horse cytochromes c were constructed with synthetic oligodeoxyribonucleotides having preferred codons and portions of the iso-1-cytochrome c-encoding gene from the yeast Saccharomyces cerevisiae. The genes were ligated into an expression vector, which contains the normal 5'- and 3'-untranslated regions of the yeast iso-1-cytochrome c gene, and were integrated in single copy into the chromosome. Yeast strains were also constructed with multiple integrated copies of the pigeon gene. The heterologous and normal mRNA levels of the single-copy strains were equivalent. Although the N-terminal methionines were completely cleaved in the heterospecific proteins, the levels of trimethylation of Lys72 and acetylation of N-terminal glycines ranged from 39-78% and 10-70%, respectively. Horse cytochrome c was produced at a nearly normal level, whereas the pigeon and tuna cytochromes c were produced at approx. 40% of the normal levels. The levels of the cytochromes c and growth of the mutant yeast strains indicated that the heterospecific cytochromes c had approx. 50% specific activity in vivo. 相似文献
27.
The role of the epidermal growth factor-1 and hydrophobic stack domains of human factor IX in binding to endothelial cells. 总被引:6,自引:0,他引:6
W F Cheung D L Straight K J Smith S W Lin H R Roberts D W Stafford 《The Journal of biological chemistry》1991,266(14):8797-8800
To determine the function and specificity in factor IX of the first epidermal growth factor (EGF)-like domain and the eight-amino acid hydrophobic stack encoded by exon C (residues 39-46), these domains were replaced by the corresponding polypeptide regions of factor X and chimeric proteins were produced in human embryo kidney cells. Both chimeras were activated by factor XIa at a rate similar to plasma factor IX and exhibited calcium-dependent fluorescence quenching similar to plasma factor IX. Both chimeras competed equally for binding to the endothelial cell receptor. Our findings make it unlikely that the first EGF-like domain or the hydrophobic stack of factor IX are responsible for the specific binding of factor IX to its endothelial cell receptor. 相似文献
28.
J. I. Moreno M. Seigelchifer J. Zorzópulos 《World journal of microbiology & biotechnology》1991,7(3):316-323
A Trypanosoma cruzi antigen gene was closed into a fusion vector based on the IgG binding domain of Staphylococcus aureus protein A. This vector transformed into Escherichia coli or Staphylococcus aureus and produced about 12 mg fusion protein/l culture. In E. coli, the product remained intracellular while in S. aureus it was excreted into the growth medium. The hybrid protein was purified by IgG Sepharose affinity chromatography. The presence of a cleavage site for enterokinase between protein A and the T. cruzi antigen in the fusion protein allowed the efficient release of the unfused antigen by enzymatic treatment. Further affinity chromatography through IgG Sepharose resulted in the production of the T. cruzi antigen free of protein A.The authors are with the Department of Molecular Genetics, BioSidus S.A., Constitución 4234, 1254, Buenos Aires, Argentina. 相似文献
29.
30.
D F Morrison P J O'Brien D R Pepperberg 《The Journal of biological chemistry》1991,266(30):20118-20123
Rhodopsin, the photosensitive protein found in rod photoreceptors, has two covalently attached palmitates that are thought to anchor a portion of the C terminus to the disc membrane, forming a fourth cytoplasmic loop. Using hydroxylamine (NH2OH) to cleave the thioester linkage, we have characterized the effect of depalmitylation on certain functional properties of rhodopsin. Treatment of rod outer segment membranes (prepared from rat retinas previously labeled in vivo with [3H]palmitate) with 1 M NH2OH typically removed greater than or equal to 75% of the [3H]palmitate initially bound to rhodopsin. Spectrophotometry of rod outer segment membranes that had been treated with 1 M NH2OH indicated preservation of 85% of the native rhodopsin and no effect on the shape of the absorbance spectrum of rhodopsin. In vivo labeled rhodopsin that had been treated with 1 M NH2OH did not reincorporate free endogenous [3H] palmitate over a 2-h incubation period. Both NH2OH-treated and untreated rhodopsin incorporated [14C]palmitate from exogenously added [14C]palmitoyl-CoA. This incorporation was substantially greater in the NH2OH-treated sample. The removal of palmitate by NH2OH inhibited rhodopsin regeneration by 44% and increased the ability of rhodopsin to activate transducin's light-dependent GTPase activity by 61%. However, the removal of palmitate from rhodopsin did not affect the light-dependent binding of transducin (T alpha and T beta gamma). 相似文献