全文获取类型
收费全文 | 13393篇 |
免费 | 830篇 |
国内免费 | 1220篇 |
专业分类
15443篇 |
出版年
2024年 | 16篇 |
2023年 | 150篇 |
2022年 | 181篇 |
2021年 | 219篇 |
2020年 | 322篇 |
2019年 | 441篇 |
2018年 | 387篇 |
2017年 | 360篇 |
2016年 | 370篇 |
2015年 | 389篇 |
2014年 | 876篇 |
2013年 | 1138篇 |
2012年 | 792篇 |
2011年 | 811篇 |
2010年 | 683篇 |
2009年 | 781篇 |
2008年 | 851篇 |
2007年 | 800篇 |
2006年 | 758篇 |
2005年 | 724篇 |
2004年 | 579篇 |
2003年 | 573篇 |
2002年 | 443篇 |
2001年 | 370篇 |
2000年 | 308篇 |
1999年 | 315篇 |
1998年 | 290篇 |
1997年 | 237篇 |
1996年 | 245篇 |
1995年 | 228篇 |
1994年 | 214篇 |
1993年 | 125篇 |
1992年 | 120篇 |
1991年 | 58篇 |
1990年 | 28篇 |
1989年 | 30篇 |
1988年 | 19篇 |
1987年 | 24篇 |
1986年 | 23篇 |
1985年 | 30篇 |
1984年 | 14篇 |
1983年 | 10篇 |
1982年 | 23篇 |
1981年 | 9篇 |
1980年 | 18篇 |
1979年 | 9篇 |
1978年 | 12篇 |
1977年 | 9篇 |
1976年 | 7篇 |
1975年 | 8篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
991.
Miyake M Sugano K Kawashima K Ichikawa H Hirabayashi K Kodama T Fujimoto H Kakizoe T Kanai Y Fujimoto K Hirao Y 《Biochemical and biophysical research communications》2007,362(4):865-871
Somatic mutations of the fibroblast growth factor receptor 3 (FGFR3) gene were detected by peptide nucleic acid (PNA)-mediated real-time PCR clamping. Mutation was detected in negative control containing only wild-type DNA due to a misincorporation of dNTPs to PNA binding sites when the amount of template DNA was decreased to 1 ng. Thus, the amount of template DNA was critical determinant of the assay sensitivity in PNA-mediated PCR clamping. Assay conditions were optimized to detect FGFR3 mutations in exons 7, 10, and 15, at a concentration of more than 1% mutated DNA using 50 ng of genomic DNA as the template. Mutations were detected in 12 of 13 (92.3%) tumor tissues and 11 of 13 (84.6%) urine samples from patients with superficial bladder cancer, while no mutations were detected in tissues and/or urine samples from patients with muscle-invasive bladder cancer or chronic cystitis. 相似文献
992.
993.
A previous genome-wide association study (GWAS) failed to discover any nucleotide sequence variant associated with susceptibility to vascular dementia (VaD) and remained a problem of false negatives produced by a low statistical power. The current study was conducted to identify such potential false negatives and to provide comprehensive evidence for the most plausible predisposing genetic factor using large-scale Korean cohorts. We identified the gene encoding retinitis pigmentosa GTPase regulator-interacting protein 1-like (RPGRIP1L) with multiple nucleotide variants associated with susceptibility to VaD by a modest significant threshold (P<10(-4)). Genetic associations were intensively examined with its sequence variants using 207 VaD patients and 207 age- and gender-matched control subjects. Genetic association analysis with dense variants in the region associated with VaD revealed 3 variants (P<0.0017) in strong linkage. Further analysis with VaD-related phenotypes using Korean Association REsource (KARE) cohort data showed that the region of the gene was associated with alanine aminotransferase (ALT), aspartate aminotransferase (AST), and blood pressure (BP) (P<7.6×10(-4)). The current study provided the first evidence of the association between RPGRIP1L gene and susceptibility of VaD. Functional studies are needed to understand underlying biological mechanism of the genetic association. 相似文献
994.
Umair Mahmood Muhammad Imran Salma Iqbal Naik Huma Arshad Cheema Anjum Saeed Muhammad Arshad Saqib Mahmood 《Gene》2012
Type I galactosemia is an inborn error resulting from mutations on both alleles of the GALT gene, which leads to the absence or deficiency of galactose-1-phosphate uridyltranseferase (GALT), the second of three enzymes catalyzing the conversion of galactose into glucose. On the basis of residual GALT activity, Type I galactosemia is classified into severe “Classical” and mild “Duarte” phenotypes. Classical galactosemia is frequently associated with S135L, Q188R and K285N mutations in the GALT gene. The functionally neutral N314D variation in the GALT gene is associated with Duarte galactosemia and is widespread among various worldwide populations. The present study aimed at detecting S135L, Q188R and K285N mutations and the N314D variant in the GALT gene by PCR using amplification refractory mutation system (ARMS). ARMS assays were established using standard DNA samples and were used for 8 galactosemia patients and 190 unrelated normal subjects all of Pakistani origin. S135L and K285N mutations were present neither in galactosemia patients nor in normal subjects. Only one galactosemia patient carried Q188R mutation that was in homozygous state. However, the N314D variant was frequently found both in affected (7 out of 16 alleles) and normal subjects (55 out of 380 alleles). This finding indicates that Duarte allele D314 might be far more common in Pakistani population than in European and North American ones. 相似文献
995.
利用RNA干扰技术降低REV3基因在人类结肠癌细胞(SW480)中的表达, 以荧光实时定量PCR检测REV3表达量的降低情况, 选择低表达效率具有统计学意义的细胞作为实验组细胞。运用细胞生长曲线、MTT、微核和姐妹染色单体交换等方法, 对实验组和对照组细胞进行细胞生长周期、增殖变化情况和遗传信息表达等指标的检测。结果显示: REV3低表达的结肠癌实验组细胞在细胞增殖以及细胞的微核和姐妹染色单体交换等遗传信息表达均明显低于结肠癌对照组细胞, 实验结果具有统计学意义(P<0.05); 结肠癌的两对照组间(阴性和空白)的结果虽然有一定的差异, 但没有统计学意义。研究结果提示, REV3低表达时, 可能对结肠癌细胞(SW480)的生长与增殖产生影响, 并对微核和姐妹染色单体交换等遗传不稳定现象的产生有一定的抑制作用。 相似文献
996.
以南瓜金辉一号(Cucurbita moschata' Jinhui1')为实验材料,利用根癌农杆菌(Agrobacterium tumefaciens)介导转化南瓜子叶节,研究了预培养时间、侵染时间、乙酰丁香酮(AS)浓度和共培养时间,抗生素羧苄青霉素(Carb)、头孢霉素(Cef)以及筛选剂卡那霉素(Kan)等因素对离体不定芽的影响,建立了南瓜最适遗传转化体系。结果表明:外植体预培养0天,侵染时间30分钟,AS浓度为100mg·L^-1,共培养5天可获得最高遗传转化效率;最适除菌剂为Cef,其最适浓度为500mg·L^-1;最适Kan筛选浓度为100mg·L^-1;在MS培养基上培养抗性芽生根,经PCR和Southern blot检测,证明为转基因植株。 相似文献
997.
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
998.
利用正交设计优化异色瓢虫SRAP-PCR反应体系 总被引:6,自引:0,他引:6
利用正交设计L16(45)对异色瓢虫Harmonia axyridis(Pallas)SRAP-PCR反应体系的5个因素(Taq酶、Mg2+、模板DNA、dNTPs、引物)在4个水平上进行优化试验,试验结果用DPS和MINITAB软件进行分析,建立了异色瓢虫SRAP-PCR反应的最佳体系,即在20μL体系中模板50~100ng、引物0.25μmol/L、dNTPs0.1mmol/L、Taq DNA聚合酶1.5U、Mg2+0.375~0.625mmol/L。并对反应体系进行梯度退火试验,得到最佳退火温度为50.3℃。这一优化系统的建立,为今后利用SRAP技术进行瓢虫遗传图谱的构建、多态性分析和基因定位奠定了技术基础。 相似文献
999.
Liang A Sha J Lu W Chen M Li L Jin D Yan Y Wang J Ping S Zhang W Wang Y Lin M 《Biotechnology letters》2008,30(8):1397-1401
A novel class II 5-enoylpyruvylshikimate-3-phosphate synthase (EPSPS) was identified from Pseudomonas stutzeri A1501 by complementation of an Escherichia coli auxotrophic aroA mutant. The single amino acid substitution of serine (Ser) for asparagine (Asn)-130 of the A1501 EPSPS enhanced resistance to 200 mM glyphosate. The mutated EPSPS had a 2.5-fold increase for IC(50) [glyphosate] value, a 2-fold increase for K (i) [glyphosate] value, but a K (m) [PEP] value similar to that of wild type. The effect of the single residue mutation on glyphosate resistance was also analyzed using a computer-based three-dimensional model. 相似文献
1000.