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91.
XU Jia-yuan LIAO Gui-cheng HU Hui-peng CHEN Xiao-zhen LIU Li-dan WU Song-ming WEN Yuan-hui LIU Yuan WENG Ya-biao LIN Rui-qing 《中国畜牧兽医》2014,41(12):30-33
In order to study whether the internal transcribed spacers (ITS) sequence could be used as a molecular marker for the species identification of rabbit coccidian, the rDNA ITS of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were amplified by polymerase chain reaction (PCR), and were cloned into pGEM-T Easy vector subsequently. The positive recombinant plasmids were identified by PCR and then sequenced. By sequence comparison and comparative analysis with the relative sequences of rabbit Eimeria spp. available in GenBank, the results showed that the lengths of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were 1065, 1009 and 1047 bp, respectively, and the sequence homologies with the same species sequences were 99.2%, 99.0% and 94.5%, respectively, while were 55.3% to 82.1% compared with corresponding sequences of other different species sequences. The phylogenetic analysis using software Mega 5.0 showed that all rabbit coccidia clustered together in a clade, which was divided into two sister lineages, corresponding to the presence or absence of oocyst residuum. The result demonstrated ITS could be used as a molecular marker for the species identification of rabbit coccidia. 相似文献
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水稻矮缩病毒(Rice dwarf virus,RDV)为双层壳、二十面体的双链RNA病毒,是水稻的重要病原物之一.文中比较了RDV中国福建分离物与日本分离物基因组各片段核苷酸序列的相似性,发现RDV两个不同分离物基因组相应片段的相似性均在92%以上,最高可达96%;分析了相应基因组片段编码的结构蛋白P1、P2、P3、P5、P7、P8及以前一直被认为是非结构蛋白的P9和非结构蛋白Pns4、Pns6、Pns10 、Pns11和Pns12在RDV复制、组装过程中的功能;回顾了RDV粒子三维结构的研究概况,对RDV粒子内、外层衣壳的晶体学结构作了较详细的描述;简要介绍了RDV复制与组装的机制,发现核心蛋白与dsRNA间相互作用成为一个单元是病毒RNA的分拣及包装到病毒粒子核心内的可能机制.同时,指出基于PDR的植物抗病毒基因工程在水稻矮缩病毒防治上的可能性. 相似文献
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为了探索人心果(Manilkara zapota L.)的转录组及奇可胶合成相关的基因,使用Illumina测序平台,对人心果果实、树皮和叶片分别进行转录组测序,使用Trinity等软件进行De novo组装和注释以及计算表达量,采用实时荧光定量PCR对转录组数据进行验证。经过组装得到162 455条unigene,总长度达139 792 553 nt,平均长度达861 nt,N50达1 544 nt。通过与NR、NT、Swiss-Prot、KEGG、COG和GO等数据库比对,共计89 628条unigene得到注释。以组装出来的unigene作为参考序列,在人心果转录组中共鉴别出57 362个SSR位点,果实、叶片和树皮表达的基因中分别检测到99 925、65 989和129109个SNP位点。在人心果转录组中共鉴别出105个与奇可胶合成相关的unigene,参与甲羟戊酸(MVA)途径的基因在果实和树皮中表达量较高,磷酸盐(MEP)途径中的基因则在叶片中的表达量较高,实时荧光定量PCR结果与转录组计算得到的表达量一致。初步推测MVA途径可能是奇可胶合成的主要途径。 相似文献
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LIU An-min LUO Ming LI Guo CAI Wang-qing LI Fang-cheng DENG Yue-fei POON Wai-sang 《园艺学报》2008,24(12):2450-2454
AIM: To construct a lentiviral vector encoding human hepatocyte growth factor (hHGF) for transfection,and to observe the expression of hHGF in human umbilical cord mesenchymal stem cells.METHODS: pUC-SRα/hHGF was subcloned into the expression vector pWPI to construct recombinant pWPI-hHGF.hHGF was identified by gene sequence.Recombinant lentivirus was produced by pWPI-hHGF,pAX2 and pMD2G altogether transient transfection into 293T cells using calcium phosphate method.The pWPI-hHGF and the contructed pWPI-GFP were transfected into human umbilical cord mesenchymal stem cells by the Lipofectamin 2000.Through counting by the fluorescent microscope,the efficiency of the transfection was identified.The expressions of hHGF and GFP in human umbilical cord mesenchymal stem cells were also detected.The concentration of hHGF in cell culture medium was determined by enzyme linked immunosorbent assay (ELISA).RESULTS: DNA sequence showed that hHGF cDNA was correctly inserted into pWPI vector.The positive rate of hHGF transfecting 293T cells was 100 %.Bright green fluorescence in the transfected cells was observed under the fluorescent microscope after 24 h transfection with lentiviral plasmid pWPI-hHGF-GFP,and the transfection rate reached 80%.The difference was distinct between the pWPI-hHGF group and control group in the secretive level of hHGF by Western blotting and the ELISA (P<0.01).CONCLUSION: The recombinant pWPI-hHGF plasmid was successfully constructed and efficient,stable and ectopic expression of hHGF was accomplished in human umbilical cord mesenchymal stem cells. 相似文献
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LUO Ming-hao HU Shu-peng WANG Rui-yu Li Chang Lü Ding-yi YANG Xi-yang LUO Su-xin 《园艺学报》2000,36(10):1739-1744
AIM To investigate whether interleukin-1β (IL-1β) regulates endothelial nitric oxide synthase (eNOS) phosphorylation at Ser1177 site in human umbilical vein endothelial cells (HUVECs), and to explore its possible mechanism. METHODS The HUVECs were randomly divided into normal control group, tumor necrosis factor-α (TNF-α) group, IL-1β group, IL-6 group, SC79 [protein kinase B (PKB/AKT) specific agonist] group and SC79+IL-1β group. Western blot was used to determine the protein levels of eNOS, p-eNOS-Ser1177, AKT and p-AKT-Ser473 in the HUVECs. Chemical colorimetry was used to detect the nitric oxide (NO) content in the culture medium of HUVECs. RESULTS No statistically significant difference of p-eNOS-Ser1177 level in HUVECs treated with TNF-α and IL-6 was observed as compared with normal control group (P >0.05), while the protein level of p-eNOS-Ser1177 in the HUVECs and the content of NO in the culture medium of HUVECs decreased significantly in IL-1β group (P <0.05), and the protein level of p-AKT-Ser473 in the HUVECs was decreased as compared with normal control group (P <0.05). The AKT agonist SC79 blocked the down-regulation effect of IL-1β on p-eNOS-Ser1177 level in the HUVECs and NO content in the culture medium of HUVECs (P< 0.05). CONCLUSION IL-1β down-regulates the protein level of p-eNOS-Ser1177 in HUVECs and affects the activity of eNOS, which may be involved in AKT/eNOS signaling pathway. 相似文献