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51.
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A. Schots J. De Boer A. Schouten J. Roosien J. F. Zil Verentant H. Pomp L. Bouwman-Smits H. Overmars F. J. Gommers B. Visser W. J. Stiekema J. Bakker 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):183-191
Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (VL) and heavy chain (VH) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location. 相似文献
53.
AIM: To investigate the contribution of angiotensin-converting enzyme inhibitor (ACEI) to the regulation of calpain system in infarcted myocardium. METHODS: Rat myocardial infarction (MI) model was established by permanent ligation of the left coronary artery. The treatment with the ACEI inhibitor rampril (1 mg·kg-1·d-1) was started 7 days prior to surgery. On day 1, 3, 7 and 14 after MI, protein levels of calpainⅠ, Ⅱ and calpastatin were determined in left ventricular free wall (LVFW), interventricular septum (IS) and right ventricule. RESULTS: CalpainⅠprotein level was increased in IS 14 d post MI, whereas the protein level of calpainⅡ was maximally increased in LVFW 3 d post MI. Rampril decreased protein up-regulation of calpainⅠ and Ⅱ, and reduced infarct size and interstitial fibrosis. Calpastatin protein expression was not affected by ACEI. CONCLUSIONS: CalpainⅠ is involved in cardiac remodelling in the late and calpainⅡ contributes to cardiac tissue damage in the early phase of MI. The heart protective effect of ACEI may be related to the inhibition of calpain system in the pathogenesis of myocardial infarction. 相似文献
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不同生态型大白菜抽薹时内源激素含量比较 总被引:5,自引:0,他引:5
大白菜〔Brassicacampestrissubspeciespekinesis(Lour. )Olsson〕先期抽薹一直困扰着春大白菜和高纬度、高海拔地区的大白菜生产。为育成冬性较强不易抽薹的品种,大白菜晚抽薹特性及利用的研究逐渐成为新的热点。不同生态型的大白菜品种,在相同的环境条件下冬性越强越不易抽薹。国外对控制或减轻未熟抽薹的环境条件有较多报道〔1~3〕,本研究主要探讨了低温处理后不同生态型的大白菜品种抽薹时内源生长素类 (IAA、IPA)、赤霉素类(GA3、GA4 )、细胞分裂素类 (ZR、DHZR)和脱落酸(ABA)含量差异,以期为选育具有较强冬性的大白菜品… 相似文献
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野生朱红硫磺菌驯化栽培研究 总被引:2,自引:2,他引:2
营养试验、菌丝体培养和出菇温度试验、培养基酸碱度试验、出菇条件试验结果表明:朱红硫磺菌菌丝体适宜生长的温度范围为15-32℃,最适温度范围为22~28℃;子实体发生的最适温度为16~25℃,气温低于16℃,子实体色泽为淡橙色,16℃以上,子实体呈鲜橙红色;培养料适宜营养C/N为20-26/1;朱红硫磺菌菌丝体适宜酸碱度范围为pH3.0~8.0,最适酸碱度为pH4.0—6.5;朱红硫磺菌的产量与光照条件无关,但光照强度影响子实体的发生时间和色泽;朱红硫磺菌栽培原料广泛,凡栽培木腐菌的培养料均可用于栽培朱红硫磺菌。 相似文献
57.
研究了苹果基因组中Tyl—copia类逆转座子的转录活性、多样性及其甲基化水平。在培养基MS+2,4-D2mg/L+BA0.2mg/L+NAA0.5mg/L上诱导了1个月的愈伤组织中用RT—PCR方法检测到了270bp的目的条带,而对照、继代组培苗和不含2,4-D的愈伤中均没有目的条带的扩增,说明一定浓度的2,4-D可以诱导苹果中Tyl—copia类逆转座子的表达活性。将目的条带回收、克隆和测序后进行分析:所克隆的21条序列虽都为Tyl-copia类逆转座子逆转录酶保守序列,但存在很大的异质性,21个克隆的核甘酸序列变化范围为196—290bp;翻译成氨基酸后存在缺失、插入、移框和终止密码子突变,7个克隆在氨基酸保守序列‘SLYGLKQ’处有1—2氨基酸位点的突变,说明这些序列对应的逆转录酶多数可能已失去了其原有功能。将21条序列进行聚类分析,除ART20外,其它克隆可聚为3类,并分别与GenBank上某些物种的相应序列同源性很高,表明在不同物种间可能存在逆转座子的横向传递作用。甲基化试验表明苹果基因组内Tyl-copia类逆转座子甲基化水平高。这些都表明Tyl-copia类逆转座子对苹果基因型的多样性及基因组的进化具有重要意义。 相似文献
58.
柑橘裂皮病类病毒的一步RT-PCR法检测及其在甜橙体内的分布 总被引:3,自引:0,他引:3
柑橘裂皮病是由柑橘裂皮病类病毒(Citrusexocortisviroid,CEVd)引起的一种重要的柑橘病害。分别采用快速微量核酸提取法和SDS-KAc抽提法在表现症状的Etrog香橼中提取核酸,采用一步RT-PCR法对CEVd进行检测,结果显示SDS-KAc法可以消除带毒样品中非特异性条带。从嫁接接毒的铜水72-1锦橙植株的接穗部取嫩叶、老叶、皮,从砧木部茎杆上取老皮以及根皮分别提取总核酸进行RT-PCR检测,分析CEVd在甜橙体内的分布情况。初步检测结果表明在接穗的嫩叶、老叶、皮,砧木部茎杆的老皮和根皮上都可以检测到CEVd,以接穗部叶和皮检出稳定性高。 相似文献
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Comparison of morphology, viability, and function between blood and milk neutrophils from peak lactating goats
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Sui Zhi Tian Chai Ju Chang Chih Chi Chiang Huo Cheng Peh Mu Chiou Huang Jai-Wei Lee Xin Zhao 《Canadian journal of veterinary research》2005,69(1):39-45
The morphological features of blood and milk neutrophils from peak lactating goats were compared using light microscopy, scanning electron microscopy and flow cytometry in order to investigate the cytological changes of neutrophils after migration into the mammary gland. The kinetics of reactive oxygen intermediates (ROI) generation and gelatinase release of blood and milk neutrophils, with or without stimulation of phorbol 12-myristate, 13-acetate ester (PMA), were used to characterize their responses to inflammatory stimuli. Neutrophils isolated from goat milk were highly segmented and contained multi-lobed nuclei. Ultrastructurally, milk neutrophils were more ruffled on the surface compared to blood neutrophils. Approximately 30% of milk neutrophils were undergoing cell death, either necrosis or apoptosis, in contrast to 8% of blood neutrophils. The ROI production of activated milk neutrophils peaked earlier than blood neutrophils, but the duration and the intensity were much less. Neutrophils from both sources augmented the release of gelatinase in response to PMA (1 ng/mL). However, the amount of gelatinase released from milk neutrophils was lower (P < 0.05) than that of blood neutrophils. In summary, more neutrophils become apoptotic and necrotic in the mammary gland, presumably due to spontaneous aging, the process of diapedesis, and the interaction with milk components. Milk neutrophils have impaired functionalities in comparison with blood neutrophils. The information is relevant when studying mammary gland immunity and related diseases, such as mastitis. 相似文献