全文获取类型
收费全文 | 4329篇 |
免费 | 146篇 |
国内免费 | 237篇 |
出版年
2023年 | 11篇 |
2022年 | 17篇 |
2021年 | 40篇 |
2020年 | 33篇 |
2019年 | 44篇 |
2018年 | 61篇 |
2017年 | 33篇 |
2016年 | 49篇 |
2015年 | 63篇 |
2014年 | 192篇 |
2013年 | 252篇 |
2012年 | 188篇 |
2011年 | 201篇 |
2010年 | 153篇 |
2009年 | 233篇 |
2008年 | 245篇 |
2007年 | 281篇 |
2006年 | 280篇 |
2005年 | 296篇 |
2004年 | 186篇 |
2003年 | 175篇 |
2002年 | 109篇 |
2001年 | 80篇 |
2000年 | 64篇 |
1999年 | 72篇 |
1998年 | 69篇 |
1997年 | 52篇 |
1996年 | 61篇 |
1995年 | 55篇 |
1994年 | 67篇 |
1993年 | 51篇 |
1992年 | 50篇 |
1991年 | 34篇 |
1990年 | 55篇 |
1989年 | 38篇 |
1988年 | 28篇 |
1987年 | 39篇 |
1986年 | 32篇 |
1985年 | 86篇 |
1984年 | 151篇 |
1983年 | 107篇 |
1982年 | 83篇 |
1981年 | 75篇 |
1980年 | 51篇 |
1979年 | 47篇 |
1978年 | 48篇 |
1977年 | 23篇 |
1976年 | 16篇 |
1975年 | 12篇 |
1973年 | 12篇 |
排序方式: 共有4712条查询结果,搜索用时 17 毫秒
71.
A combined physical and genetic map of theCorynebacterium glutamicum ATCC 13032 chromosome was constructed using pulsed-field gel electrophoresis (PFGE) and hybridizations with cloned gene probes. Total genomic DNA was digested with the meganucleasesSwaI (5-ATTTAAAT-3),PacI (5-TTAATTAA-3), andPmeI (5-GTTTAAAC-3) yielding 26, 27, and 23 fragments, respectively. The chromosomal restriction fragments were then separated by PFGE. By summing up the lengths of the fragments generated with each of the three enzymes, a genome size of 3082 +/- 20 kb was determined. To identify adjacentSwaI fragments, a genomic cosmid library ofC. glutamicum was screened for chromosomal inserts containingSwaI sites. Southern blots of the PFGE gels were hybridized with these linking clones to connect theSwaI fragments in their natural order. By this method, about 90% of the genome could be ordered into three contigs. Two of the remaining gaps were closed by cross-hybridization of blottedSwaI digests using as probesPacI andPmeI fragments isolated from PFGE gels. The last gap in the chromosomal map was closed by hybridization experiments using partialSwaI digestions, thereby proving the circularity of the chromosome. By hybridization of gene probes toSwaI fragments separated by PFGE about 30 genes, including rRNA operons, IS element and transposon insertions were localized on the physical map. 相似文献
72.
Radioadaptive response: Efficient repair of radiation-induced DNA damage in adapted cells 总被引:13,自引:0,他引:13
To verify the hypothesis that the induction of a novel, efficient repair mechanism for chromosomal DNA breaks may be involved in the radioadaptive response, the repair kinetics of DNA damage has been studied in cultured Chinese hamster V79 cells with single-cell gel electrophoresis. The cells were adapted by priming exposure with 5 cGy of γ-rays and 4-h incubation at 37°C. There were no indication of any difference in the initial yields of DNA double-strand breaks induced by challenging doses from non-adapted cells and from adapted cells. The rejoining of DNA double-strand breaks was monitored over 120 min after the adapted cells were challenged with 5 or 1.5 Gy, doses at the same level to those used in the cytogenetical adaptive response. The rate of DNA damage repair in adapted cells was higher than that in non-adapted cells, and the residual damage was less in adapted cells than in non-adapted cells. These results indicate that the radioadaptive response may result from the induction of a novel, efficient DNA repair mechanism which leads to less residual damage, but not from the induction of protective functions that reduce the initial DNA damage. 相似文献
73.
Hisao Kurazono Shinji Yamasaki Orn-anong Ratchtrachenchai G. Balakrish Nair Yoshifumi Takeda 《Microbiology and immunology》1996,40(4):303-305
Vibrio cholerae O139 isolated from different countries, as well as from different locations within a country, were examined using macrorestriction DNA analysis to determine the clonality of the O139 strains. NotI digests of genomic DNA of representative strains from Nepal, India, Bangladesh, China, Thailand, and Malaysia revealed very similar but not identical patterns. Examinations of the banding patterns generated by pulsed-field gel electrophoresis of strains isolated within countries revealed complete homogeneity. These results further reiterate the spread of an identical clone of V. cholerae O139 although it appears that genetic polymorphism among the O139 strains is becoming apparent. 相似文献
74.
将1mm厚凝固于复印膜上的水琼脂(15%~2.0%)凝胶板侵入含12mmol/L植酸钠的Gly—HCl缓冲液中达1h以上,取出干至胶表面无水迹,于上加Aspergillussp.59—2植酸酶或与电泳后的凝胶板紧贴10~60min。然后水平置于恒温水浴锅中反应一定时间,取出浸入1mol/LH2SO4-2%(NH4)6Mn7O24-10%FeSO4相似文献
75.
Joaquin Royo Isabel Diaz Pablo Rodriquez-Palenzuela Pilar Carbonero 《Plant molecular biology》1996,31(5):1051-1059
The geneItr1, encoding trypsin inhibitor BTI-CMe, has been obtained from a genomic library ofHordeum vulgare L. The gene has no introns and presents in its 5-upstream region 605 bp that are homologous to the long terminal repeats (LTR) of the copia-like retro-transposon Bare-1. Functional analysis of theItr1 promoter by transient expression in protoplasts derived from different barley tissues, has shown that in this system theItr1 promoter retains its endosperm specifity and thetrans-regulation mediated by theLys3a gene. The proximal promoter extending 343 bp upstream of the translation initiation ATG codon is sufficient to confer fullGUS expression and for endosperm specifity. In protoplasts derived from thelys3a mutant, Risø 1508,GUS activity was less than 5% of that obtained with the same constructs in the protoplasts of wild-type Bomi from which it derives. Gel retardation experiments, after incubation with proteins obtained from both types of endosperm nuclei, also show differential patterns. Possible reasons for these differences are discussed.Equal authours 相似文献
76.
77.
复合淀粉凝胶电泳同工酶分析 总被引:2,自引:0,他引:2
为了克服水解马铃薯淀粉不易获得的困难,并使“I发片淀粉凝胶电泳同工酶分析”更容易开展,普通的化学试剂马铃薯淀粉(或精制食用马铃薯淀粉)和可溶性淀粉混合物加入适当 剂被用来代替水解马铃薯淀粉制作凝胶。试验结果表明:用8 ̄10%的上述混合淀粉(5:3),添加1%的琼脂粉和2 ̄4%的蔗糖,所制成的“复合淀粉凝胶”可以很好地被切片,并成功地对许多不同类群的植物材料的PGM、PGI、MDH、AAT、SKDH 相似文献
78.
Chieko Takaya Ayako Kosaka Kyouko Kohno Toshihisa Kusano Ken-Ichi Nakamura 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1995,112(4):727-732
Tetrahymena thermophila could still swim after incubation of the cell body at 40°C for 30 min, whereas Tetrahymena pyriformis did not show any motility after the treatment. Turbidity measurements revealed that axonemes of T. pyriformis lost ATP-dependent sliding activity by the heat treatment, whereas those of T. thermophilia still had the activity under the same conditions. In connection with this difference in susceptibility to high temperature, the biochemical characteristics of dyneins were compared between the two species of Tetrahymena. Axonemal dyneins from the two species had significant vanadate-sensitive ATPase activity even after the heat treatment. Native gel electrophoresis and the following two-dimensional electrophoresis showed that the outer arm dynein of T. thermophilia is more stable in maintaining native configuration than that of T. pyriformis against the heat treatment, although both treated dyneins keep three (α, β and γ) subunits. Analysis by peptide mapping demonstrated that β- and γ-subunits of the outer arm dynein are considerably different in amino acid sequences between the two species. These results imply that dynein of T. thermophilia changed their amino acid sequences and biochemical characteristics to adapt to high temperature. 相似文献
79.
You-Di Liao 《Molecular biology reports》1994,20(3):149-154
Ribonucleases are widely found in the tissues of living organisms, but the functions of individual ribonucleases are not clear. To facilitate characterization of individual ribonucleases, I have developed a rapid method to separate and identify each ribonuclease from a crude sample by gel electrophoresis instead of by time-consuming purification steps. The ribonucleases in a crude sample are first separated by RNA-cast SDS-polyacrylamide gel electrophoresis and then eluted from the gel after ethidium bromide staining. To determine the base specificity of each ribonuclease, a 5 labelled oligonucleotide with known sequence is added to the enzyme eluate and the digested products are analyzed by denaturing gel electrophoresis. The base specificity of bovine pancreatic ribonuclease (RNase A), bullfrog oocyte-specific ribonuclease (RC-RNase), human serum ribonucleases and sweet potato leaf ribonucleases were determined by this method. Other properties of individual ribonucleases, e.g. substrate preference, may also be determined from crude samples by this method without further purification steps.Abbreviations RNase
ribonuclease
- SDS
sodium dodecyl sulfate 相似文献
80.
Jang LK 《Biotechnology and bioengineering》1994,43(2):183-185
The diffusivity of Cu(2+) in calcium alginate beads calculated by the shrinking core model (SCM) was reevaluated in this work. The results obtained in this work were significantly different than those by the original authors. There were excellent agreements between the results obtained by the SCM in this work and those by the more rigorous linear absorption model (LAM) by the original authors. (c) 1994 John Wiley & Sons, Inc. 相似文献