全文获取类型
收费全文 | 3484篇 |
免费 | 129篇 |
国内免费 | 462篇 |
专业分类
林业 | 73篇 |
农学 | 139篇 |
基础科学 | 20篇 |
182篇 | |
综合类 | 1164篇 |
农作物 | 108篇 |
水产渔业 | 312篇 |
畜牧兽医 | 1866篇 |
园艺 | 56篇 |
植物保护 | 155篇 |
出版年
2024年 | 18篇 |
2023年 | 40篇 |
2022年 | 131篇 |
2021年 | 132篇 |
2020年 | 118篇 |
2019年 | 148篇 |
2018年 | 86篇 |
2017年 | 138篇 |
2016年 | 163篇 |
2015年 | 135篇 |
2014年 | 191篇 |
2013年 | 180篇 |
2012年 | 275篇 |
2011年 | 300篇 |
2010年 | 205篇 |
2009年 | 215篇 |
2008年 | 165篇 |
2007年 | 226篇 |
2006年 | 218篇 |
2005年 | 142篇 |
2004年 | 109篇 |
2003年 | 90篇 |
2002年 | 69篇 |
2001年 | 77篇 |
2000年 | 71篇 |
1999年 | 44篇 |
1998年 | 52篇 |
1997年 | 38篇 |
1996年 | 41篇 |
1995年 | 35篇 |
1994年 | 31篇 |
1993年 | 26篇 |
1992年 | 25篇 |
1991年 | 20篇 |
1990年 | 26篇 |
1989年 | 24篇 |
1988年 | 21篇 |
1987年 | 11篇 |
1986年 | 8篇 |
1984年 | 3篇 |
1983年 | 3篇 |
1982年 | 2篇 |
1981年 | 3篇 |
1980年 | 6篇 |
1979年 | 3篇 |
1974年 | 1篇 |
1973年 | 2篇 |
1962年 | 1篇 |
1956年 | 6篇 |
1955年 | 1篇 |
排序方式: 共有4075条查询结果,搜索用时 15 毫秒
41.
为了制备草鱼重组TAB1蛋白(rCiTAB1)及其特异性抗体,首先以草鱼头肾组织c DNA为模板,PCR扩增Ci TAB1基因全长序列,并依次构建重组克隆质粒p MD19-T-Ci TAB1与重组表达质粒pET-32a-Ci TAB1。该重组表达质粒经0.5 mmol·L~(-1) IPTG,37℃诱导表达12 h后获得以包涵体形式表达的重组CiTAB1蛋白(rCiTAB1)。然后,采用3种不同方法对包涵体蛋白进行变性,发现与高浓度尿素直接变性法和洗涤后尿素变性法相比,洗涤后梯度尿素变性法处理后的蛋白纯度最好,经透析复性后得到浓度为2 mg·mL~(-1)的r Ci TAB1。最后,将rCiTAB1蛋白与白油佐剂及免疫增强剂混合,室温下混合1.5h乳化成免疫原,3次免疫新西兰大白兔制备兔抗CiTAB1抗体,经间接ELISA方法和免疫琼脂双向扩散试验测得免疫后第33天血清中特异性抗体的效价分别为1∶1 048 576和1∶16。Western blot检测到一条分子量约为72 kDa的特异性条带,表明该抗体能特异性识别r Ci TAB1蛋白。研究结果为后续深入研究CiTAB1蛋白功能提供了物质基础。 相似文献
42.
43.
J R López S Núñez B Magariños N Castro J I Navas R de la Herran A E Toranzo 《Journal of fish diseases》2009,32(7):603-610
The first isolation of Tenacibaculum maritimum from wedge sole, Dicologoglossa cuneata, is reported. The pathogen was recovered from ulcers of cultured fish, from three different outbreaks. The six isolates obtained were biochemically and serologically characterized and diagnosis was confirmed by polymerase chain reaction using specific primers and partial 16S rRNA gene sequencing. The isolates constituted a homogeneous phenotypic group; however, they belong to two of the different serotypes described within this species. A virulence evaluation of the isolates using Wedge sole fry was also performed. 相似文献
44.
Wandong Fu Yan Wang Bin Zheng Miaofei Liao Weijie Zhang 《Journal Of Aquatic Food Product Technology》2013,22(3):270-280
Pepsin-soluble collagen (PSC) was isolated from Peru squid (Dosidicus gigas) skin and physicochemical properties of the PSC were determined. The PSC exhibited a maximum absorbance at 220 nm. Sodium dodecyl sulfate polyacrylamide gel electrophoresis suggested the collagen containing α1 and α2 chain was classified as type I collagen. Amino acid composition indicated that the collagen had lower amino acid content than that of mammalian collagen. Denaturation temperature (Td) of the PSC was 26.8°C. The PSC had relatively high solubility in alkaline condition or NaCl concentrations below 2%. Fourier transform infrared spectroscopy investigations showed the existence of helical arrangements of collagen. The lyophilized collagen had a uniform and regular network structure. These results suggested that Peru squid skin was a potential source of collagen for further research and application. 相似文献
45.
An aquabirnavirus (ABV) and a formalin-inactivated betanodavirus [redspotted grouper nervous necrosis virus (RGNNV)] were investigated for their potential to prevent RGNNV-induced viral nervous necrosis (VNN) in the sevenband grouper, Epinephelus septemfasciatus (Thunberg). Three groups of fish were injected intramuscularly with ABV, intraperitoneally with inactivated RGNNV (iRGNNV) or with both ABV and iRGNNV. At 3, 7, 14, 21 and 28 days post-injection (p.i.), fish were challenged by intramuscular injection of RGNNV. Control fish, which received neither ABV nor iRGNNV, showed high mortalities in all RGNNV challenges. Fish that received only ABV exhibited relative percent survival (RPS) of >60 against RGNNV challenges at 3, 7, 14 and 21 days p.i., but not at 28 days p.i., while fish that received only iRGNNV showed significantly higher protection against RGNNV challenges only at 21 and 28 days p.i. In contrast, fish that received both ABV and iRGNNV showed 60 or higher RPS against all RGNNV challenges. Fish inoculated with iRGNNV with or without ABV exhibited similar high titres of neutralizing antibodies to RGNNV at 14, 21 and 28 days p.i. These results indicate that combined inoculation with iRGNNV and ABV conferred both rapid non-specific and delayed specific protection against VNN. 相似文献
46.
Cyanobacteria, particularly thermophilic strains, represent an important potential source of EPSs, harboring structural complexity that predicts diverse and specific bioactive potential. The thermophilic cyanobacteria Gloeocapsa gelatinosa, isolated from a natural hot source in Ain Echfa (Tunisia), was cultivated in a cylindrical reactor, and the production of biomass and EPSs was investigated. Results revealed that the strain is amongst the most efficient EPSs producers (0.89 g L−1) and that EPSs production was not correlated with the growth phase. EPSs were sulfated heteropolysaccharides containing carbohydrates (70%) based on nine different monosaccharides, mainly mannose (22%), and with the presence of two uronic acids. EPSs were formed by two polymers moieties with a molecular weight of 598.3 ± 7.2 and 67.2 ± 4.4 kDa. They are thermostable in temperatures exceeding 100 °C and have an anionic nature (zeta potential of −40 ± 2 mV). Atomic force microscopy showed that EPSs formed multimodal lumps with 88 nm maximum height. EPSs presented high water holding capacity (70.29 ± 2.36%) and solubility index (97.43 ± 1.24%), and a strong bivalent metal sorption capacity especially for Cu2+ (91.20 ± 1.25%) and Fe2+ (75.51 ± 0.71%). The antioxidant activity of G. gelatinosa EPSs was investigated using four methods: the β-carotene-bleaching activity, DPPH assays, iron-reducing activity, and metal-chelating activity. EPS has shown high potential as free radicals’ scavenger, with an IC50 on DPPH (0.2 g L−1) three-fold lower than ascorbic acid (0.6 g L −1) and as a metal chelating activity (IC50 = 0.4 g L−1) significantly lower than EDTA. The obtained results allow further exploration of the thermophilic G. gelatinosa for several biotechnological and industrial applications. 相似文献
47.
Guoyi Niu Dilok Wongsathein Sukolrat Boonyayatra Rutch Khattiya 《Aquaculture Research》2019,50(12):3643-3652
Edwardsiella tarda is a pathogen that causes edwardsiellosis in aquatic animals. The emergence of multiple antibiotic‐resistant strains makes antibiotic treatment difficult. This study aimed to investigate the antibiotic susceptibility patterns and the genotypic characterization of E. tarda isolated from cage‐cultured red tilapia in Thailand. A total of 30 isolates were identified as E. tarda using biochemical and molecular analysis. The disc diffusion method for testing antibiotic susceptibility showed all the isolates were resistant to colistin sulphate and oxolinic acid. High levels of resistance to amoxicillin, ampicillin, ceftazidime, oxytetracycline and sulphamethoxazole/trimethoprim were observed as well. The multiple antibiotic resistance index ranged from 0.25 to 0.92, indicating that these isolates had been exposed to high risk sources of contamination where antibiotics were commonly used. All the isolates carried the blaTEM gene based on polymerase chain reaction (PCR). The tetA and sul3 genes were detected in 90% (27/30) and 26.7% (8/30) of the isolates respectively. Nine different genetic groups of isolates were obtained using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC‐PCR). A correlation between genetic types and multiple antibiotic‐resistant patterns was found. These results highlight the potential risks of multiple antibiotic‐resistant isolates for humans and the environment. 相似文献
48.
49.
为从蛋白质水平研究细胞因子在鲤体内免疫应答过程中的合成变化,本研究采用PCR技术克隆TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β基因含有部分抗原决定簇的片段,引入双酶切位点Bam HⅠ和HindⅢ后连接至p ET-32a/21a,构建相应的表达载体,制备多克隆抗体。采用ELISA检测抗体效价,并以此作为实验工具,检测经嗜水气单胞菌感染后鲤血清中炎性细胞因子的合成变化。结果显示,基因TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β融合蛋白分子量分别约为31.8、31.7、35.3、32.5、18.0和33.6 ku;抗体效价达到2.4×106;在病原菌感染后的不同阶段,促炎细胞因子TNF-α、IL-1β、IL-6、IL-12和抗炎细胞因子IL-10、TGF-β呈现出不同的合成变化。研究表明,制备的抗体具有较高的效价、亲和力和特异性,可用于鲤细胞因子的定量研究,该抗体的获得为鲤免疫应答与细胞因子合成的系统研究奠定了基础。同时,获取的鲤细胞因子TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β的抗体亦可用于其他鱼类细胞因子蛋白质水平的定量研究。 相似文献
50.
草鱼干扰素的分离纯化及某些理化和生物学特性 总被引:3,自引:0,他引:3
通过DEAE-Sepharose阴离子交换层析、Sephacryl S-200凝胶层析、高效液相层析和梯度聚丙烯酰张胶电泳等技术,对病毒诱生的草鱼血清干扰素进行了分离纯化。纯化的草鱼干扰素在SDS-聚丙烯酰胺凝胶电泳中呈现单一组份,分子量为38kD,等电点为5.25,过碘酸-Schiff试剂反应表现为典型的糖蛋白染色特征。理化和生物学特性研究表明,草鱼干扰素具有100000g(2h)离心不沉降、耐 相似文献