全文获取类型
收费全文 | 3534篇 |
免费 | 152篇 |
国内免费 | 365篇 |
出版年
2024年 | 2篇 |
2023年 | 39篇 |
2022年 | 48篇 |
2021年 | 77篇 |
2020年 | 80篇 |
2019年 | 90篇 |
2018年 | 77篇 |
2017年 | 67篇 |
2016年 | 72篇 |
2015年 | 91篇 |
2014年 | 141篇 |
2013年 | 199篇 |
2012年 | 118篇 |
2011年 | 185篇 |
2010年 | 118篇 |
2009年 | 183篇 |
2008年 | 224篇 |
2007年 | 214篇 |
2006年 | 223篇 |
2005年 | 207篇 |
2004年 | 187篇 |
2003年 | 189篇 |
2002年 | 157篇 |
2001年 | 122篇 |
2000年 | 89篇 |
1999年 | 99篇 |
1998年 | 73篇 |
1997年 | 84篇 |
1996年 | 78篇 |
1995年 | 70篇 |
1994年 | 76篇 |
1993年 | 64篇 |
1992年 | 52篇 |
1991年 | 46篇 |
1990年 | 37篇 |
1989年 | 30篇 |
1988年 | 16篇 |
1987年 | 12篇 |
1986年 | 10篇 |
1985年 | 26篇 |
1984年 | 17篇 |
1983年 | 12篇 |
1982年 | 17篇 |
1981年 | 5篇 |
1980年 | 7篇 |
1979年 | 9篇 |
1978年 | 3篇 |
1977年 | 4篇 |
1974年 | 2篇 |
1973年 | 3篇 |
排序方式: 共有4051条查询结果,搜索用时 921 毫秒
41.
Foreign protein expression from the commonly used SV40 promoter has been found to be primarily during the S-phase of the cell cycle. Simple mathematical models with this cell cycle phase dependent expression of foreign protein suggest that the specific production rate will be proportional to the cell growth rate, which is particularly disadvantageous in high cell density fed-batch or perfusion bioreactors. In this study we investigate this predicted relationship between the production rate and growth rate by culturing recombinant CHO cells in a continuous suspension bioreactor. One CHO cell line, GS-26, has been stably transfected with the plasmid pSVgal, which contains the E. coli lac Z gene under the control of the SV40 promoter. This GS-26 cell line was grown in suspension cultures over a range of specific growth rates in batch and continuous modes. The intracellular -galactosidase activity was assayed using a standard spectrophotometric method after breaking the cells open and releasing the enzyme. A strong growth associated relationship is found between the intracellular -galactosidase content and the specific growth rate in batch and continuous cultures, as predicted. 相似文献
42.
Tohru Tsukui Sanae Miyake Sadahiro Azuma Hirotake Ichise Izumu Saito Yutaka Toyoda 《Molecular reproduction and development》1995,42(3):291-297
Replication-defective recombinant adenovirus, Adex4SRLacZL, was used as a vector for transferring exogenous genes in mouse zona pellucida-free eggs at the pronuclear stage. The vector contained the E. coli LacZ reporter gene under the control of the SRα promoter (SV40 early promoter-fused HTLV-I LTR), and the expression of the reporter gene was examined during preimplantation development in culture. Histochemical staining of the embryos for β-galactosidase activity showed that the exogenous LacZ gene as expressed in 98% of the embryos at the morula-blastocyst stages. As in the microinjection method, the exogenous genes could be pursued from the 2-cell stage. Neither apparent morphological changes nor cytotoxic effects were observed. Both the percentages of embryos expressing reporter genes and the rate of development to the blastocyst stage were higher in the adenovirus vector-treated embryos than in the microinjected ones. These results suggest that the adenovirus vector system is a useful tool in investigating the genetic control of early mammalian development. © 1995 wiley-Liss, Inc. 相似文献
43.
Structural organization and differential expression of three stilbene synthase genes located on a 13 kb grapevine DNA fragment 总被引:10,自引:0,他引:10
Wilfried Wiese Barbara Vornam Elvira Krause Helmut Kindl 《Plant molecular biology》1994,26(2):667-677
A 13 kb DNA fragment was isolated from a grapevine (Vitis var. Optima) genomic library by hybridizing with elicitor-induced stilbene synthase cDNA as a probe. After fragmentation with Eco RI, subcloning and sequencing, two full-size stilbene synthase genes (Vst1 and Vst2) and the 3 end of a third stilbene synthase gene (Vst3) were located within the 13 kb fragment. Vst1 and Vst2, differing only slightly in the coding region, are distinguished in the intron size and in the structure of the promoter region. The 5 flanking region of gene Vst1 contains a TATAA box at nucleotide –48. The substantial structural differences found for the promoters of the two genes are paralleled by a striking difference in the expression of the two genes in elicitor-treated cells. Moreover, the accumulation upon elicitation of six different stilbene synthase mRNAs was studied and found to differ by two orders of magnitude. 相似文献
44.
A. J. Soitamo G. Zhou A. K. Clarke G. Öquist E-M. Aro P. Gustafsson 《Plant molecular biology》1994,26(2):709-721
The unicellular cyanobacterium Synechococcus sp. PCC 7942 has three psbA genes encoding two different forms of the photosystem II reaction centre protein D1 (D1:1 and D1:2). The level of expression of these psbA genes and the synthesis of D1:1 and D1:2 are strongly regulated under varying light conditions. In order to better understand the regulatory mechanisms underlying these processes, we have constructed a strain of Synechococcus sp. PCC 7942 capable of over-producing psbA mRNA and D1 protein. In this study, we describe the over-expression of D1:1 using a tac-hybrid promoter in front of the psbAI gene in combination with lacI
Q
repressor system. Over-production of D1:1 was induced by growing cells for 12 h at 50 mol photons m-2 s-1 in the presence of 40 or 80 g/ml IPTG. The amount of psbAI mRNA and that of D1:1 protein in cells grown with IPTG was three times and two times higher, respectively. A higher concentration of IPTG (i.e., 150 g/ml) did not further increase the production of the psbAI message or D1:1. The over-production of D1:1 caused a decrease in the level of D1:2 synthesised, resulting in most PSII reaction centres containing D1:1. However, the over-production of D1:1 had no effect on the pigment composition (chlorophyll a or phycocyanin/number of cells) or the light-saturated rate of photosynthesis. This and the fact that the total amounts of D1 and D2 proteins were not affected by IPTG suggest that the number of PSII centres within the membranes remained unchanged. From these results, we conclude that expression of psbAI can be regulated by using the tac promoter and lacI
Q
system. However, the accumulation of D1:1 protein into the membrane is regulated by the number of PSII centres. 相似文献
45.
46.
A genomic clone for the cyc07 gene, which is expressed specifically at the S phase during the cell cycle in synchronous cultures of periwinkle (Catharanthus roseus) cells, was isolated. Determination of the nucleotide sequence of the clone revealed that the cyc07 gene consists of seven exons separated by six introns. Genomic Southern analysis indicated that the cyc07 gene is present as a single copy per haploid genome in periwinkle. Expression of related genes was detected in a wide range of other plants. Transgenic Arabidopsis plants were generated that expressed the gene for -glucuronidase (GUS) under the control of the promoter of the cyc07 gene. The tissue-specific pattern of expression directed by the promoter was investigated by analysis of GUS activity. Histochemical tests demonstrated that 589 bp of the 5-upstream sequence of the cyc07 gene could direct specifical expression of the GUS reporter gene in meristematic tissues in transgenic plants. The spatial pattern of expression directed by the promoter was closely correlated with meristematic activity and cell proliferation, suggesting an association between the function of the cyc07 gene and cell proliferation. 相似文献
47.
Arsenio Fernández-López Carlos Soria Victoria Revilla Tomás Gómez Pedro Calvo 《Neurochemical research》1994,19(4):445-449
At 4 days after the implantation of two subcutaneous 75 mg morphine pellets in the back skin, rats were morphine-dependent. In the three layers studied in the occipital cortex we found that the values of the 2-adrenergic agonist [3H]bromoxidine binding increased with respect to animals implanted with placebo pellets. Typical behavioral and physiological symptoms of the abstinence syndrome appeared 30 minutes after administration of naloxone, [3H]bromoxidine binding values being similar to those obtained in animals implanted with placebo pellets. The pattern of response of the [3H]bromoxidine binding was similar in the hippocampus and the superficial gray layer of the superior colliculus of the mesencephalon, but the differences were not statistically significant in these areas. This paper concludes that exist brain regional differences in the 2-adrenoreceptors response under morphine-treatment and possibly under naloxone-induced morphine abstinence syndrome. 相似文献
48.
We have constructed a luc reporter vector for Dictyostelium discoideum using a 626-bp fragment from the nuclear-associated plasmid Ddp2. The ori from Ddp2 is localized within this fragment and was used to provide an autonomous replication sequence for the reporter vector. This reporter vector was stably retained in D. discoideum AX3K cells without alteration. The vector molecule was also found to exist in relatively low copy number compared to other Dictyostelium vectors in the transformed cells. We demonstrated the utility of this vector as a reporter vector with glycogen synthase promoter/luc fusions of varying sizes. 相似文献
49.
50.