全文获取类型
收费全文 | 15921篇 |
免费 | 961篇 |
国内免费 | 1509篇 |
专业分类
林业 | 1344篇 |
农学 | 1018篇 |
基础科学 | 792篇 |
1608篇 | |
综合类 | 7403篇 |
农作物 | 1021篇 |
水产渔业 | 681篇 |
畜牧兽医 | 2585篇 |
园艺 | 1051篇 |
植物保护 | 888篇 |
出版年
2024年 | 117篇 |
2023年 | 306篇 |
2022年 | 683篇 |
2021年 | 761篇 |
2020年 | 694篇 |
2019年 | 671篇 |
2018年 | 474篇 |
2017年 | 726篇 |
2016年 | 531篇 |
2015年 | 744篇 |
2014年 | 819篇 |
2013年 | 936篇 |
2012年 | 1339篇 |
2011年 | 1296篇 |
2010年 | 1274篇 |
2009年 | 1083篇 |
2008年 | 1193篇 |
2007年 | 1035篇 |
2006年 | 911篇 |
2005年 | 652篇 |
2004年 | 436篇 |
2003年 | 294篇 |
2002年 | 337篇 |
2001年 | 368篇 |
2000年 | 252篇 |
1999年 | 136篇 |
1998年 | 62篇 |
1997年 | 42篇 |
1996年 | 33篇 |
1995年 | 31篇 |
1994年 | 25篇 |
1993年 | 15篇 |
1992年 | 24篇 |
1991年 | 18篇 |
1990年 | 15篇 |
1989年 | 7篇 |
1988年 | 7篇 |
1987年 | 6篇 |
1986年 | 7篇 |
1985年 | 2篇 |
1984年 | 2篇 |
1983年 | 2篇 |
1982年 | 3篇 |
1981年 | 4篇 |
1979年 | 3篇 |
1965年 | 1篇 |
1962年 | 1篇 |
1956年 | 8篇 |
1955年 | 2篇 |
1920年 | 1篇 |
排序方式: 共有10000条查询结果,搜索用时 696 毫秒
121.
为探究肌生长抑制素(myostatin,MSTN)基因对牛肌肉发育的具体调控机制,本研究选取同一牛场健康鲁西黄牛10头,其中通过转基因技术得到的基因编辑牛MSTN-/-和同种非转基因野生型牛各5头。分别采集两组牛腿臀肌肉样品,利用IIlumina HiSeq高通量测序技术进行转录组测序分析,通过生物信息学方法比较两组样本间的差异表达基因,并进行GO和KEGG富集分析,最后利用实时荧光定量PCR验证转录组测序数据。结果显示,基因编辑型牛和野生型牛之间共检测到18 071个基因。在log2|FoldChange|≥ 1.48条件下,筛选出406个差异表达基因,其中347个显著上调,59个显著下调。GO功能富集分析显示,MSTN基因编辑后显著影响915个功能类别(P<0.05),差异基因主要参与结合、生物系统调节、免疫系统等相关功能。KEGG通路富集分析结果共涉及211个通路,差异基因主要富集在细胞黏附分子、趋化因子信号通路、细胞因子互作等信号通路上,进一步从中筛选出可能参与细胞生长、肌肉发育的差异基因(CD14、KIT、CSF1R、FBP1、DUSP4、ULBP21、PRKCB、SPN、CHAD、SRC)。实时荧光定量PCR检测结果显示,所选差异基因表达水平与转录组表达水平一致,证明测序结果的可靠性。本研究结果表明,MSTN基因发挥作用后可以介导多个下游基因表达,从而影响相关信号通路及生物学过程;同时,所筛选出的差异表达基因可作为进一步研究骨骼肌调控机制的候选靶标。 相似文献
122.
为探究肌肉生长抑制素(myostatin,MSTN)对牛骨骼肌生长发育的作用机制,本研究前期利用定量蛋白质组学与磷酸化蛋白质组学分析野生型蒙古牛(MG.WT)和MSTN+/-蒙古牛(MG.MSTN+/-)腿臀肌肌肉组织中蛋白质水平和磷酸化修饰水平的差异变化,使用已建立的牛骨骼肌卫星细胞体外诱导成肌分化模型,检测设计合成的MSTN siRNA (si-MSTN)干扰效果;采用实时荧光定量PCR和Western blotting方法检测转染si-MSTN的增殖期(GM)和分化第3天(DM3)牛骨骼肌卫星细胞中肌动蛋白细胞骨架调节通路相关基因的mRNA和蛋白水平的表达变化,研究敲低MSTN表达对肌动蛋白细胞骨架调节通路的影响。结果显示,在MSTN+/-蒙古牛肌肉组织中共鉴定到16个肌动蛋白细胞骨架调节通路相关基因表达丰度上调;转染si-MSTN细胞中的MSTN表达水平极显著降低(P<0.01);在转染si-MSTN的GM期牛骨骼肌卫星细胞中,肌动蛋白细胞骨架调节通路相关基因ENAH、ACTN4和Cdc42的mRNA水平均显著升高(P<0.05),PFN1、RhoA和ACTN4的蛋白水平均显著或极显著升高(P<0.05;P<0.01);在转染si-MSTN的DM3牛骨骼肌卫星细胞中,ENAH、CFL1、SCIN和Cdc42基因mRNA水平均显著升高(P<0.05),RhoA基因mRNA水平极显著升高(P<0.01),PFN1和ACTN4的蛋白水平均显著升高(P<0.05)。结果表明,干扰MSTN可以促进肌动蛋白细胞骨架调节通路相关基因的表达,探明了MSTN可能通过介导肌动蛋白细胞骨架调节通路影响牛骨骼肌卫星细胞增殖和成肌分化的分子机制,为进一步研究MSTN对牛成肌分化的调控机制提供参考。 相似文献
123.
124.
125.
ZHANG Hai ZHOU Bi-jun YANG Zhong-cheng LIAO Mei WANG Kai-gong WEN Ming CHENG Zhen-tao WANG Wei FENG Xu-fang 《中国畜牧兽医》2016,43(11):2834-2843
In order to study and analyze L1 gene of bovine papillomavirus(BPV)in Guizhou province,the L1 gene of BPV-GZ01 strain was amplified,cloned and sequenced using bioinformatic softwares and methods,and the secondary structure,tertiary structure,B-cell preponderant epitope,conserved domains analysis, transmembrane domain and signal peptide of L1 gene were predicted.The results showed that the length of L1 gene was 1 494 bp,encoding 497 amino acids.The L1 gene of BPV-GZ01 strain shared an amino acid identities of 98.6%,99.4%,98.4%,94.4% and 91.3%,and a nucleotide identities of 99.1%,99.8%,99.4%,87.6% and 82.8% with those of BPV2,BPV2-SW01,BPV2-AKS01,BPV13 and BPV1 strains,respectively.The results of phylogenetic tree analysis indicated that there was a close relationship between BPV-GZ01 and BPV2-SW01 strains.The prediction of secondary structure of L1 protein indicated that the random coil,extended strand and alphahelix took a higher percentage.The L1 protein was supposed contain 6 potential antigen epitopes.And no transmembrane domains and no signal peptide were found.The tertiary structure of L1 protein was curved spiral structure.These results provided a theoretical basis for immunologic diagnosis and further research of nucleic acid vaccine of BPV. 相似文献
126.
YUE Yuan CHEN Hong-yan WANG Jia-wei XU Ming-qiang DING Yu JIANG Hao GAO Yan ZHANG Jia-bao YAN Shou-qing 《中国畜牧兽医》2016,43(3):585-591
The aim of this study was to investigate the differential expression genes induced by ApoCⅢ,and study the function of ApoCⅢ.Porcine aortic vascular endothelial cells were successfully isolated using enzyme digestion,and then screened the differential expression genes induced by ApoCⅢ using the Solexa high-throughput sequencing technology.The results showed 647 differential expression genes,including 390 up-regulated genes and 257 down-regulated genes.The qRT-PCR results verified that the gene expression results from Solexa sequencing data were reliable.GO and Pathway analysis showed that the function of differential expression genes were related to immune response,cell apoptosis and death.These findings suggested that ApoCⅢ affected the physiological function of porcine aortic endothelial cells by the molecular pathways of inflammation,cell adhesion and apoptosis,which provided a theoretical basis for further understanding the molecular mechanisms of atherosclerosis caused by ApoCⅢ. 相似文献
127.
In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×105 anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×104 anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV. 相似文献
128.
WANG Lei LV Yang ZHANG Yue XIE Jia-qian ZHAO Di YAN Li-qiong MEI Hui-min WANG Lei YI Dan CHEN Hong-bo DING Bin-ying HOU Yong-qing WU Tao 《中国畜牧兽医》2016,43(5):1355-1360
The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro. 相似文献
129.
针对现有日光温室人工通风方式劳动强度大、室内气温分布不均匀等问题,在温室内安装2台或3台智能型屋脊通风设备对温室进行2段或3段智能通风,并在北京地区冬季日光温室与人工通风控制进行了对比。结果表明:在保温被揭开期间,当采用3段智能通风时,温室内空气温度最大值与最小值之差为(1.1±0.5)℃;而2段智能控制和人工控制分别为(2.3±1.1)℃和(3.8±1.3)℃。此外,应用该智能屋脊通风技术,还能将667m2均效益提高0.97万元。因此,智能型屋脊通风设备可显著改善温室内温度分布的均匀性,提高种植的经济收益。 相似文献
130.