全文获取类型
收费全文 | 765篇 |
免费 | 82篇 |
国内免费 | 142篇 |
出版年
2024年 | 5篇 |
2023年 | 19篇 |
2022年 | 55篇 |
2021年 | 74篇 |
2020年 | 46篇 |
2019年 | 53篇 |
2018年 | 47篇 |
2017年 | 38篇 |
2016年 | 49篇 |
2015年 | 45篇 |
2014年 | 58篇 |
2013年 | 50篇 |
2012年 | 63篇 |
2011年 | 62篇 |
2010年 | 28篇 |
2009年 | 23篇 |
2008年 | 28篇 |
2007年 | 33篇 |
2006年 | 36篇 |
2005年 | 38篇 |
2004年 | 20篇 |
2003年 | 23篇 |
2002年 | 18篇 |
2001年 | 7篇 |
2000年 | 3篇 |
1999年 | 5篇 |
1998年 | 9篇 |
1997年 | 3篇 |
1996年 | 7篇 |
1995年 | 5篇 |
1994年 | 7篇 |
1993年 | 5篇 |
1991年 | 3篇 |
1990年 | 1篇 |
1989年 | 3篇 |
1988年 | 2篇 |
1987年 | 4篇 |
1986年 | 4篇 |
1985年 | 3篇 |
1974年 | 3篇 |
1973年 | 1篇 |
1972年 | 2篇 |
1971年 | 1篇 |
排序方式: 共有989条查询结果,搜索用时 468 毫秒
51.
Purification of chloroplast cytochrome b 559 总被引:1,自引:0,他引:1
H S Garewal J Singh A R Wasserman 《Biochemical and biophysical research communications》1971,44(6):1300-1305
52.
Bacterial populations common to healthy human guts may play important roles in human health. A new strategy for discovering genomic sequences as markers for these bacteria was developed using Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR fingerprinting. Structural features within microbial communities are compared with ERIC-PCR followed by DNA hybridization to identify genomic fragments shared by samples from healthy human individuals. ERIC-PCR profiles of fecal samples from 12 diseased or healthy human and piglet subjects demonstrated stable, unique banding patterns for each individual tested. Sequence homology of DNA fragments in bands of identical size was examined between samples by hybridization under high stringency conditions with DIG-labeled ERIC-PCR products derived from the fecal sample of one healthy child. Comparative analysis of the hybridization profiles with the original agarose fingerprints identified three predominant bands as signatures for populations associated with healthy human guts with sizes of 500, 800 and 1000 bp. Clone library profiling of the three bands produced 17 genome fragments, three of which showed high similarity only with regions of the Bacteroides thetaiotaomicron genome, while the remainder were orphan sequences. Association of these sequences with healthy guts was validated by sequence-selective PCR experiments, which showed that a single fragment was present in all 32 healthy humans and 13 healthy piglets tested. Two fragments were present in the healthy human group and in 18 children with non-infectious diarrhea but not in eight children with infectious diarrhea. Genome fragments identified with this novel strategy may be used as genome-specific markers for dynamic monitoring and sequence-guided isolation of functionally important bacterial populations in complex communities such as human gut microflora. 相似文献
53.
The chromosome constitution of hybrids and chromatin patterns of Agropyron elongatum (Host)Neviski in F5 somatic hybrid lines -1–3 and I-1-9 between Triticum aestivum L. and A. elongatum were analyzed. Based on the statistic data
of pollen mother cells, F5 I-1-9 and-1-3 had 20–21 bivalents with a frequency of 84.66% and 85.28%, of which, 89.83% and 89.57% were ring bivalents.
The result indicated that both hybrid lines were basically stable in the chromosome constitution and behavior. RAPD analysis
showed that the two hybrids contained biparental and integrated DNA. GISH (Genome in situ hybridization) revealed that in
the form of small chromosome segments, A. elongatum chromatin was scattered on 4–6 wheat chromosomes near by the region of
centromere and telomere in the two hybrid lines. SSR analysis indicated that A. elongatum DNA segments were distributed on
the 2A, 5B, 6B and 2D wheat chromosomes in the hybrids, which was in accordance with the GISH results that small-segments
intercalated poly-site. 相似文献
54.
Most bacteria produce the dUMP precursor for thymine nucleotide biosynthesis using two enzymes: a dCTP deaminase catalyzes the formation of dUTP and a dUTP diphosphatase catalyzes pyrophosphate release. Although these two hydrolytic enzymes appear to catalyze very different reactions, they are encoded by homologous genes. The hyperthermophilic archaeon Methanococcus jannaschii has two members of this gene family. One gene, at locus MJ1102, encodes a dUTP diphosphatase, which can scavenge deoxyuridine nucleotides that inhibit archaeal DNA polymerases. The second gene, at locus MJ0430, encodes a novel dCTP deaminase that releases dUMP, ammonia, and pyrophosphate. Therefore this enzyme can singly catalyze both steps in dUMP biosynthesis, precluding the formation of free, mutagenic dUTP. Besides differing from the previously characterized Salmonella typhimurium dCTP deaminase in its reaction products, this archaeal enzyme has a higher affinity for dCTP and its steady-state turnover is faster than the bacterial enzyme. Kinetic studies suggest: 1) the archaeal enzyme specifically recognizes dCTP; 2) dCTP deamination and dUTP diphosphatase activities occur independently at the same active site, and 3) both activities depend on Mg(2+). The bifunctional activity of this M. jannaschii enzyme illustrates the evolution of a suprafamily of related enzymes that catalyze mechanistically distinct reactions. 相似文献
55.
The protein product of the Methanococcus jannaschii MJ0768 gene has been expressed in Escherichia coli, purified to homogeneity, and shown to catalyze the GTP-dependent addition of two l-glutamates to the l-lactyl phosphodiester of 7,8-didemethyl-8-hydroxy-5-deazariboflavin (F(420)-0) to form F(420)-0-glutamyl-glutamate (F(420)-2). Since the reaction is the fifth step in the biosynthesis of coenzyme F(420), the enzyme has been designated as CofE, the product of the cofE gene. Gel filtration chromatography indicates CofE is a dimer. The enzyme has no recognized sequence similarity to any previously characterized proteins. The enzyme has an absolute requirement for a divalent metal ion and a monovalent cation. Among the metal ions tested, a mixture of Mn(2+), Mg(2+), and K(+) is the most effective. CofE catalyzes amide bond formation with the cleavage of GTP to GDP and inorganic phosphate, likely involving the activation of the free carboxylate group of F(420)-0 to give an acyl phosphate intermediate. Evidence for the occurrence of this intermediate is presented. A reaction mechanism for the enzyme is proposed and compared with other members of the ADP-forming amide bond ligase family. 相似文献
56.
The lyotropic liquid-crystalline behavior of trimethylsilyl hydroxypropylcellulose (TMS-HPC) is reported in this paper. The introduction of the trimethylsilyl (TMS) group in the parent HPC increases the solubility in organic solvents, and the lyotropic mesophase can be formed in concentrated acetone solution. The critical concentration (C*) in acetone is approximately 36%. The liquid crystalline nature of TMS-HPC/acetone solution was confirmed by PLM, and the mechanism of liquid crystallization was studied by FTIR and WAXD methods. 相似文献
57.
The enzyme responsible for observed IMP cyclohydrolase activity in Methanococcus jannaschii was purified and sequenced: its genetic locus was found to correspond to gene MJ0626. The MJ0626 gene was cloned, and its protein product was expressed in Escherichia coli and shown to catalyze the cyclization of 5-formylamidoimidazole-4-carboxamide ribonucleotide to IMP. The enzyme has no sequence similarity to known enzymes, and its catalytic properties appear distinct from any characterized IMP cyclohydrolase. The purO gene for the enzyme is currently found only in the domain ARCHAEA: 相似文献
58.
The protein product of the Methanococcus jannaschii MJ1256 gene has been expressed in Escherichia coli, purified to homogeneity, and shown to be involved in coenzyme F(420) biosynthesis. The protein catalyzes the transfer of the 2-phospholactate moiety from lactyl (2) diphospho-(5')guanosine (LPPG) to 7,8-didemethyl-8-hydroxy-5-deazariboflavin (Fo) with the formation of the L-lactyl phosphodiester of 7,8-didemethyl-8-hydroxy-5-deazariboflavin (F(420)-0) and GMP. On the basis of the reaction catalyzed, the enzyme is named LPPG:Fo 2-phospho-L-lactate transferase. Since the reaction is the fourth step in the biosynthesis of coenzyme F(420), the enzyme has been designated as CofD, the product of the cofD gene. The transferase requires Mg(2+) for activity, and the catalysis does not appear to proceed via a covalent intermediate. To a lesser extent CofD also catalyzes a number of additional reactions that include the formation of Fo-P, when the enzyme is incubated with Fo and GDP, GTP, pyrophosphate, or tripolyphosphate, and the hydrolysis of F(420)-0 to Fo. All of these side reactions can be rationalized as occurring by a common mechanism. CofD has no recognized sequence similarity to any previously characterized enzyme. 相似文献
59.
用SDS.NaClO从重组大肠杆菌中分离聚-β-羟基丁酸酯 总被引:4,自引:0,他引:4
聚β羟基丁酸酯(PHB)是微生物合成的一种以颗粒状态存在于细胞中的高分子聚合物,由于它具有生物可降解性、生物相容性等特性,在医学上具有独特而广阔的应用前景。从微生物细胞中分离PHB的方法有溶剂萃取法[1]、化学试剂法[24]和酶法[5]。目前工... 相似文献
60.
大鼠催乳素基因真核细胞可表达性质粒的构建及应用研究 总被引:4,自引:0,他引:4
735bp的PRLcDNA片段从质粒PRL-SP65#1中回收后,用粘性末端连接法将其重组到真核表达载体pcDNA3上,筛选出正向连接重组体pcDNA3-PRLS和反向连接重组体pcDNA3-PRLAS。将重组体pcDNA3-PRLs和空载体pcDNA3分别转入NIH3T3细胞系,用G418筛选出阳性细胞后与未转染的NIH3T3细胞在加E2和不加E2的情况下,用原位杂交的方法,分别用PRLcDNA探针和原癌基因c-H-rascDNA探针进行检测,未转染的NIH3T3细胞在加E2和不加E2时都几乎无催乳素基因的表达,同样,转入空载体的NIH3T3细胞也无PRL的表达,而转入重组体pcDNA3-PRLS的NIH3T3细胞则有大量的PRL基因的表达,与对照组相比有显著差异(P<0.01)。正常和转入空载体的NIH3T3细胞有一定程度的原癌基因c-H-ras的表达,当分别加入E2和转入重组体pcDNA3-PRLS后,NIH3T3细胞中的c-H-ras基因表达水平都显著升高(P<0.05)。 相似文献