全文获取类型
收费全文 | 34665篇 |
免费 | 2017篇 |
国内免费 | 5134篇 |
出版年
2024年 | 51篇 |
2023年 | 519篇 |
2022年 | 589篇 |
2021年 | 1018篇 |
2020年 | 900篇 |
2019年 | 1182篇 |
2018年 | 938篇 |
2017年 | 894篇 |
2016年 | 971篇 |
2015年 | 1181篇 |
2014年 | 1597篇 |
2013年 | 2216篇 |
2012年 | 1594篇 |
2011年 | 1618篇 |
2010年 | 1413篇 |
2009年 | 1745篇 |
2008年 | 1914篇 |
2007年 | 2059篇 |
2006年 | 2119篇 |
2005年 | 1970篇 |
2004年 | 1804篇 |
2003年 | 1744篇 |
2002年 | 1584篇 |
2001年 | 1291篇 |
2000年 | 1080篇 |
1999年 | 1032篇 |
1998年 | 874篇 |
1997年 | 757篇 |
1996年 | 736篇 |
1995年 | 714篇 |
1994年 | 680篇 |
1993年 | 482篇 |
1992年 | 417篇 |
1991年 | 353篇 |
1990年 | 310篇 |
1989年 | 221篇 |
1988年 | 235篇 |
1987年 | 220篇 |
1986年 | 164篇 |
1985年 | 150篇 |
1984年 | 127篇 |
1983年 | 68篇 |
1982年 | 88篇 |
1981年 | 48篇 |
1980年 | 45篇 |
1979年 | 33篇 |
1978年 | 30篇 |
1977年 | 9篇 |
1976年 | 17篇 |
1950年 | 6篇 |
排序方式: 共有10000条查询结果,搜索用时 31 毫秒
121.
Thomas F. Holzman Christine C. Chung Rohinton Edalji David A. Egan Earl J. Gubbins Annemarie Rueter Gail Howard Lana K. Yang Terry M. Pederson Grant A. Krafft et al. 《Journal of Protein Chemistry》1990,9(6):663-672
The gene for human preprorenin was obtained from total RNA prepared from primary human chorion cells. An expression vector was constructed containing an SV40 early promoter, a human preprorenin cDNA, bovine growth hormone poly-A addition signal, and a dihydrofolate reductase (dhfr) expression cassette. This vector was inserted into the DXB-11 Chinese hamster ovary (CHO) cell line. The recombinant protein was exported by CHO cells into the tissue culture media. At harvest the prorenin levels ranged from 1–5 mg/L. For prorenin isolation the cell culture supernatants were processed by filtration, concentration, dialysis, and batch extraction. Preparative-scale isolation of prorenin was accomplished using blue-dye chromatography and size-exclusion chromatography. The isolated prorenin yielded a single SDS-gel band with Mr 40,000. The proprotein was characterized with respect to N-terminal sequence and N-linked sugar composition. Trypsin-activated renin prepared from the proprotein was characterized with respect to N-terminal sequence andpH-activity profile. Enzyme activity was measured with a newly developed fluorogenic peptide substrate containing the P6-P3 sequence of human angiotensinogen. 相似文献
122.
The 6.5 kb HindIII DNA fragment of the Lactococcus lactis subsp. cremoris H2 plasmid pDI21 was cloned into Escherichia coli POP13 with NM1149, and also directly into Lactococcus lactis subsp. lactis 4125 using a newly-constructed broad host-range vector pFX1. Proteinase was experessed in both transformed organisms. The proteinase resembles a PI type since it preferentially degraded -casein. The restriction map of the 6.5 kb proteinase gene fragment has minor differences from those of published plamid proteinase genes. High-efficiency electroporation with pFX1 provides a direct approach for gene cloning in lactococci.Abbreviations cfu
colony forming units
- HEPES
N-[2-hydroxyethyl]piperazine-N-[2-ethanesulphonic acid]
Dedicated to Prof. Dr. G. Drews on the occasion of his 65th birthday 相似文献
123.
In the present communication, we report the identification of a new gene family which encodes the protein subunits of the proteasome. The proteasome is a high-Mr complex possessing proteolytic activity. Screening a Drosophila λgt11 cDNA expression library with the proteasome-specific antibody N19-28 we isolated a clone encoding the 28-kDa No. 1 proteasome protein subunit. In accordance with the nomenclature of proteasome subunits in Drosophila, the corresponding gene is designated PROS-28.1, and it encodes an mRNA of 1.1 kb with an open reading frame of 249 amino acids (aa). Genomic Southern-blot hybridization shows PROS-28.1 to be a member of a family of related genes. Analysis of the predicted aa sequence reveals a potential nuclear targeting signal, a potential site for tyrosine kinase and a potential cAMP/cGMP-dependent phosphorylation site. The aa sequence comparison of the products of PROS-28.1 and PROS-35 with the C2 proteasome subunit of rat shows a strong sequence similarity between the different proteasome subunits. The data suggest that at least a subset of the proteasome-encoding genes belongs to a family of related genes (PROS gene family) which may have evolved from a common ancestral PROS gene. 相似文献
124.
银合欢基因文库的构建和种子贮藏蛋白基因的分离 总被引:1,自引:0,他引:1
潘志强 《Acta Botanica Sinica》1990,32(6):420-424
本文以λEMBL_3为载体,构建了银合欢(Leucaena leucocephala)的基因文库,所得重组子为3.5×10~6pfu。以大豆种子贮藏蛋白α′亚基基因为探针,从基因文库中分离得到了4个阳性克隆,并初步绘制了其中3个重组子的物理图谱。结果表明在这3个重组子的基因内部有一致的酶切位点。亚克隆的部分核苷酸序列与 GenBank 中的基因序列比较,表明与大豆种子贮藏蛋白α′亚基基因高度同源。 相似文献
125.
抗阿特拉津转基因大豆植株后代的遗传分析 总被引:9,自引:0,他引:9
本试验用阿特拉津溶液涂抹、荧光诱导动力学检测、分子杂交等方法对抗阿特拉津转基因大豆植株的后代进行了鉴定,在第二代及第三代中检测到了抗性基因的存在,表明从龙葵中得到的此抗阿特拉津 psbA 基因不仅能导人大豆叶绿体基因组中获得表达,而且可以遗传到后代。 相似文献
126.
127.
128.
129.
Purification and properties of theDrosophila zen protein 总被引:1,自引:0,他引:1
Summary The zen protein is encoded by the zerknullt gene required for normal early development inDrosophila. Like many regulatory proteins of this type, zen contains a 60 amino acid homeobox sequence. We have purified the zen protein
and studied its solution behavior and its interaction with DNA. The zen protein exists as a monomer in solution with a molecular
weight of about 40000. It binds specifically to a site about 900 bases upstream from thezen gene. Within this binding site DNase protection experiments indicate that binding is confined to two regions approximately
11 and 14 bases in length that are separated by about 30 base pairs. The protein concentration dependence of the binding curve
suggests that protein binding is non cooperative. 相似文献
130.
Robert Sweetland Patricia C. Sheppard Janice G. Dodd Robert J. Matusik 《Molecular and cellular biochemistry》1988,84(1):3-15
Summary After castration, the rat dorsolateral prostate M-40 mRNA initially decreased then rebounded to precastrated levels. The cellular site of M-40 expression and its renewed expression after castration was defined by in situ hybridization histochemistry. In situ hybridization with either a 32P-labeled or biotin-labeled M-40 cDNA probe demonstrated that M-40 mRNA levels were higher in the lateral than dorsal prostate. A second androgen regulated gene, RWB, also was highly expressed in the lateral prostate. The biotinylated cDNA probes provided microscopic resolution of the expressing cells, revealing two distinct morphologies of lateral epithelium which expressed both the M-40 and RWB mRNA. These morphologies appeared in ducts which contained either epithelial cell sheets that were highly convoluted or thinner epithelial cells with a minimal degree of convolution. The RWB mRNA decreased in both cell populations in response to androgen withdrawal. The decline and reappearance of M-40 mRNA also appeared in both epithelial cell types. These data demonstrated that after castration the M-40 mRNA initially decreased as expected for an androgen sensitive gene and then progressed to a fully inducible state. The mechanism of this progression remains to be elucidated. 相似文献