全文获取类型
收费全文 | 8920篇 |
免费 | 385篇 |
国内免费 | 401篇 |
出版年
2023年 | 68篇 |
2022年 | 125篇 |
2021年 | 112篇 |
2020年 | 139篇 |
2019年 | 185篇 |
2018年 | 236篇 |
2017年 | 134篇 |
2016年 | 169篇 |
2015年 | 190篇 |
2014年 | 695篇 |
2013年 | 798篇 |
2012年 | 543篇 |
2011年 | 492篇 |
2010年 | 324篇 |
2009年 | 431篇 |
2008年 | 426篇 |
2007年 | 469篇 |
2006年 | 412篇 |
2005年 | 398篇 |
2004年 | 295篇 |
2003年 | 315篇 |
2002年 | 255篇 |
2001年 | 231篇 |
2000年 | 167篇 |
1999年 | 190篇 |
1998年 | 201篇 |
1997年 | 171篇 |
1996年 | 164篇 |
1995年 | 203篇 |
1994年 | 193篇 |
1993年 | 165篇 |
1992年 | 144篇 |
1991年 | 84篇 |
1990年 | 82篇 |
1989年 | 44篇 |
1988年 | 36篇 |
1987年 | 35篇 |
1986年 | 32篇 |
1985年 | 37篇 |
1984年 | 59篇 |
1983年 | 44篇 |
1982年 | 56篇 |
1981年 | 37篇 |
1980年 | 29篇 |
1979年 | 15篇 |
1978年 | 13篇 |
1977年 | 24篇 |
1976年 | 9篇 |
1975年 | 7篇 |
1973年 | 10篇 |
排序方式: 共有9706条查询结果,搜索用时 15 毫秒
41.
Polymerase chain reaction (PCR) technology has revolutionized the process of isolating and amplifying segments of DNA. One powerful application of PCR is its use in precise site-directed mutagenesis (SDM). SDM provides an elegant tool for scientists and engineers to explore biocatalytic mechanisms and processes to understand the structural-functional relationships of enzymes and other proteins. This article reviews techniques and methodology used in site-directed mutagenesis of genes by PCR. 相似文献
42.
Effects of different concentrations of active ingredient of the herbicide pyramin on metabolic activities of Fusarium solani and Sclerotium rolfsii were examined. High concentrations of this herbicide (1000 and 2000 g mL-1 for F. solani and 100 and 200 g mL-1 for S. rolfsii) had inhibitory effects on the metabolic activities of both fungi. These were demonstrated by significant decreases in growth, and increases in rates of CO2 evolved, O2 consumed and keto acids produced. These were accompanied by increased rates of sugar, nitrate and inorganic phosphorus absorption as well as lowered rates of synthesis of carbohydrates and insoluble nitrogenous (including protein) and phosphorus (including RNA-P and DNA-P) compounds. In addition, rates of excretion of both nitrogen and phosphorus fractions by the mycelial mats were increased.A concentration of 25 g mL-1 exerted little or no effect on the metabolic activities of these fungi, although S. rolfsii was somewhat sensitive to this concentration. 相似文献
43.
Universal primers for amplification of three non-coding regions of chloroplast DNA 总被引:90,自引:0,他引:90
Six primers for the amplification of three non-coding regions of chloroplast DNA via the polymerase chain reaction (PCR) have been designed. In order to find out whether these primers were universal, we used them in an attempt to amplify DNA from various plant species. The primers worked for most species tested including algae, bryophytes, pteridophytes, gymnosperms and angiosperms. The fact that they amplify chloroplast DNA non-coding regions over a wide taxonomic range means that these primers may be used to study the population biology (in supplying markers) and evolution (inter- and probably intraspecific phylogenies) of plants. 相似文献
44.
Jac M. M. J. G. Aarts Jan G. J. Hontelez Peter Fischer Ruud Verkerk Albert van Kammen Pim Zabel 《Plant molecular biology》1991,16(4):647-661
With a view to cloning the root-knot nematode resistance gene Mi in tomato by chromosome walking, we have developed a molecular probe for the tightly linked acid phosphatase-1 (Aps-1) locus. The acid phosphatase-1 allozyme (APS-11), encoded by the Aps-1
1 allele originating from Lycopersicon peruvianum, was purified to apparent homogeneity from tomato roots and suspension cells. Microsequencing of CNBr and tryptic peptides generated from APS-11 provided a partial amino acid sequence, which accounted for approximately 23% of the protein and revealed two stretches of homology with soybean proteins KSH3 and VSP27, comprising 22 matches within 26 amino acid residues. The partial amino acid sequence information enabled us to isolate a 2.4 kb genomic Aps-1
1 sequence by means of the polymerase chain reaction (PCR), primed by degenerate pools of oligodeoxyribonucleotides, synthesized on the basis of the amino acid sequences. Synthesis of the 2.4 kb PCR product was specific for genomic templates carrying the L. peruvianum Aps-1
1 allele. Crucial to the priming specificity and the synthesis of the 2.4 kb genomic sequence was the use of degenerate primer pools in which the number of different primer species was limited by incorporating deoxyinosine phosphate residues at three and four base ambiguities. In using cDNA as a template, a 490 bp sequence was obtained, indicating a high proportion of intron sequences in the 2.4 kb genomic Aps-1
1 sequence. The Aps-1
1 origin of the PCR product was confirmed by RFLP (restriction fragment length polymorphism) analysis, using both a chromosome 6 substitution line and a pair of nearly isogenic lines, differing for a small chromosomal region around the Aps-1/Mi loci. 相似文献
45.
Ignacio R. Rodriguez Pedro Gonzales J. Samuel Zigler Teresa Borrás 《生物化学与生物物理学报:疾病的分子基础》1992,1180(1):44-52
A congenital cataract present in guinea pigs provided a unique opportunity to study a hereditary lens diseases at the molecular level. ζ-crystallin, one of the most abundant guinea pig lens proteins, was found to be altered in the lens of cataractous animals. Several ζ-crystallin cDNA clones were isolated from a cataractous lens library and found to contain a 102-bp deletion towards the 3′ end of the coding region. The deletion does not interfere with the reading frame but results in a protein 34 amino acids shorter. Sequence analysis of a normal genomic ζ-crystallin clone revealed that the missing 102-bp fragment corresponds to an entire exon (exon 7). PCR analysis of the genomic DNA isolated from cataractous animals showed that exon 7, though missing from the mRNA, is intact in the cataractous genome. Further sequence analysis of the α-crystallin gene disclosed a dinucleotide delection of the universal AG at the acceptor splice-site of intron 6 of the mutant gene. The presence of this mutation results in the skipping of exon 7 during the mRNA processing which in turn results in the altered ζ-crystallin protein. This if the first time a genomic mutation in an enzyme/crytallin gene has been directly linked to a congenital cataract. 相似文献
46.
47.
The effect of ciprofibrate, a hypolipidemic drug, was examined in the metabolism of palmitic (C16:0) and lignoceric (C24:0) acids in rat liver. Ciprofibrate is a peroxisomal proliferating drug which increases the number of peroxisomes. The palmitoyl-CoA ligase activity in peroxisomes, mitochondria and microsomes from ciprofibrate treated liver was 3.2, 1.9 and 1.5-fold higher respectively and the activity for oxidation of palmitic acid in peroxisomes and mitochondria was 8.5 and 2.3-fold higher respectively. Similarly, ciprofibrate had a higher effect on the metabolism of lignoceric acid. Treatment with ciprofibrate increased lignoceroyl-CoA ligase activity in peroxisomes, mitochondria and microsomes by 5.3, 3.3 and 2.3-fold respectively and that of oxidation of lignoceric acid was increased in peroxisomes and mitochondria by 13.4 and 2.3-fold respectively. The peroxisomal rates of oxidation of palmitic acid (8.5-fold) and lignoceric acid (13.4-fold) were increased to a different degree by ciprofibrate treatment. This differential effect of ciprofibrate suggests that different enzymes may be responsible for the oxidation of fatty acids of different chain length, at least at one or more step(s) of the peroxisomal fatty acid -oxidation pathway. 相似文献
48.
Three cases of abnormal expression of the equine protease inhibitory alleles, Pi F, L, and S1, were observed following the examination of 30,000 plasma samples by one-dimensional acid (pH 4.6) polyacrylamide gel electrophoresis. Characterization of the abnormal proteins in terms of isoelectric point, molecular mass, inhibitory spectra, and sialic acid content was performed using one- and two-dimensional electrophoretic techniques. The Pi F and S1 abnormalities were postulated to be the result of amino acid substitutions causing alterations in the processing of the carbohydrate side chains. No explanation could be offered for the Pi L abnormality other than a charge shift mutation. Abnormal types, F*, L*, and S*1 behaved as alleles but the distribution of L* in offspring from one stallion (present in only 6 of 83 offspring) differed significantly from expectation.This work was supported by a grant from the Australian Stud Book, Alison Road, Randwick, N.S.W. 2031. 相似文献
49.
Hans-B?rje Jansson A. Jeyaprakash Gerald C. Coles Nahum Marban-Mendoza Bert M. Zuckerman 《Journal of nematology》1986,18(4):570-574
Caenorhabditis elegans and Panagrellus redivivus were investigated for surface carbohydrates using fluorescent-labelled and ferritin-labelled lectins. Rhodamine-labelled Concanavalin A was specifically located in the cephalic region of both species. Rhodamine-labelled wheat germ agglutinin was located over the entire cuticle of P. redivivus but was absent on C. elegans. Rhodamine-labelled peanut agglutinin and Limax flavus agglutinin did not label nematodes of either species. Galactose and sialic acid were not detected on either species, whereas mannose-glucose residues were specifically localized in the head areas of both species. No detectable N-acetylglucosamine occurred on C. elegans, but it was evenly distributed over the cuticle surface of P. redivivus. 相似文献
50.