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A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm. 相似文献
3.
Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability
to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases
and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an
intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS.
The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system.
The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365. 相似文献
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Based on biochemical assays and electrophoretical methods, the inhibitory effects of three plant meals (soybean meal, wheat meal, winged bean meal) on digestive alkaline proteases of discus were investigated. Casein assays revealed that increasing levels of soybean meal caused a linear inhibitory effect on activity of protease. SDS-PAGE images revealed that trypsin and chymotrypsin were the inhibited enzymes. Soybean showed the lowest inhibition rate followed by wheat meal and raw winged bean. There was a quadratic relationship between wheat meal levels and its inhibition of protease acitivity. The highest inhibitory effect was obtained with the winged bean meal with inhibition of caseinolytic activities ranging from 3.6–98.6%. Results from this study showed the potential of both soybean meal and wheat meal as ingredients for practical diet for discus, while demonstrating the need for further improvement in processing method for winged bean meal. 相似文献
6.
不同生长阶段斑节对虾消化酶活性变化 总被引:3,自引:0,他引:3
通过对斑节对虾不同生长阶段蛋白酶、脂肪酶及淀粉酶活性变化的分析,探讨了斑节对虾在生长过程中对蛋白质、脂肪和碳水化合物的消化利用。结果表明,根据消化酶活性,可将斑节对虾生长分为两个阶段。第1阶段,在体长10~12.5 cm阶段之前,随着个体的增大蛋白酶和脂肪酶活性不断升高;第2阶段,在12.5 cm之后,蛋白酶和脂肪酶活性呈现出一定程度的下降。肝胰腺、胃、肠3种不同消化器官的消化酶活性在不同生长期也不一致,大致上蛋白酶活性从高到低依次为:肝胰腺>胃>肠;脂肪酶活性:肠>肝胰腺>胃;淀粉酶活性:肠>胃>肝胰腺。 相似文献
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Jian-Bin Zhou Qi-Cun Zhou Shu-Yan Chi Qi-Hui Yang Chu-Wu Liu 《Aquaculture (Amsterdam, Netherlands)》2007,270(1-4):186-192
A growth experiment was conducted to determine the optimal dietary protein requirement for juvenile ivory shell reared in indoor aerated aquaria. Six isoenergetic experimental diets using fish meal, casein and gelatin as protein sources were formulated to contain graded levels of protein (27, 33, 38, 43, 49 and 54% of dry diet, respectively). Triplicate groups of 40 shells (average weight 93.50 ± 1.70 mg) were stocked in 120-l tanks and fed to apparent satiation twice daily for 8 weeks. The results showed that the growth performance and feed utilization were significantly affected by dietary protein level (P < 0.05). Maximum weight gain, mean protein gain, specific growth rate and soft body to shell ratio occurred at 43% dietary protein level (P < 0.05). There were significant differences in protein, lipid, moisture and ash content in soft body; except that ash content in shell was not significantly affected by dietary protein level. Pepsin activity in soft body tissue significantly increased with dietary protein level up to 43%, and trypsin-like enzyme activity increased with dietary protein level up to 49%. However, lipase activity in soft body decreased with increasing dietary protein level. However, no significant differences (P < 0.05) in survival, calcium, phosphorus concentration in the shell and soft body were found among dietary treatments. Quadratic regression analysis of weight gain against dietary protein level indicated that the optimal dietary protein requirement for maximum growth and feed utilization of juvenile ivory shell is 45% of dry diet. 相似文献
9.
以南极磷虾(Euphausia superb)为研究对象,通过硫酸铵分级沉淀、Phenyl-Sepharose疏水层析、DEAE-Sepharose FF离子交换层析等方法,从南极磷虾体内分离纯化出胰蛋白酶。其纯化倍数为5.44倍,比活力为38.3 U/mg,得率为26%。SDS-PAGE电泳结果显示,该酶的分子质量为28 ku。蛋白酶最适温度为37℃、最适pH为7.5,Mg2+、Ca2+、Mn2+对南极磷虾蛋白酶具有激活性,Zn2+、Cu2+、Fe3+具有酶活抑制性,其中Cu2+的抑制性最强。酶的动力学实验结果表明,以BApNA为底物测得Km为0.073 mmol/L,Vmax为1.44×10-2mmol/L·s,kcat为0.6S-1,kcat/Km为8.22×103,PMSF作为蛋白酶抑制剂,对南极磷虾蛋白酶作用机制为不可逆抑制。 相似文献
10.
A review of cyanobacterial odorous and bioactive metabolites: Impacts and management alternatives in aquaculture 总被引:2,自引:0,他引:2
Juliette L. Smith Greg L. Boyer Paul V. Zimba 《Aquaculture (Amsterdam, Netherlands)》2008,280(1-4):5-20
Increased demand has pushed extensive aquaculture towards intensively operated production systems, commonly resulting in eutrophic conditions and cyanobacterial blooms. This review summarizes those cyanobacterial secondary metabolites that can cause undesirable tastes and odors (odorous metabolites) or are biochemically active (bioactive metabolites) in marine and freshwater, extensive and intensive aquaculture systems. For the scope of this paper, biochemically active metabolites include (1) toxins that can cause mortality in aquaculture organisms or have the potential to harm consumers via accumulation in the product (hepatotoxins, cytotoxins, neurotoxins, dermatoxins, and brine shrimp/molluskal toxins), (2) metabolites that may degrade the nutritional status of aquaculture species (inhibitors of proteases and grazer deterrents) or (3) metabolites that have the potential to negatively affect the general health of aquaculture species or aquaculture laborers (dermatoxins, irritant toxins, hepatotoxins, cytotoxins). Suggestions are made as to future management practices in intensive and extensive aquaculture and the potential exposure pathways to aquaculture species and human consumers are identified. 相似文献