全文获取类型
收费全文 | 1238篇 |
免费 | 84篇 |
国内免费 | 156篇 |
出版年
2023年 | 24篇 |
2022年 | 18篇 |
2021年 | 31篇 |
2020年 | 31篇 |
2019年 | 47篇 |
2018年 | 28篇 |
2017年 | 31篇 |
2016年 | 32篇 |
2015年 | 34篇 |
2014年 | 82篇 |
2013年 | 81篇 |
2012年 | 63篇 |
2011年 | 79篇 |
2010年 | 50篇 |
2009年 | 78篇 |
2008年 | 84篇 |
2007年 | 81篇 |
2006年 | 75篇 |
2005年 | 64篇 |
2004年 | 59篇 |
2003年 | 46篇 |
2002年 | 44篇 |
2001年 | 31篇 |
2000年 | 18篇 |
1999年 | 30篇 |
1998年 | 16篇 |
1997年 | 11篇 |
1996年 | 11篇 |
1995年 | 13篇 |
1994年 | 12篇 |
1993年 | 11篇 |
1992年 | 4篇 |
1991年 | 6篇 |
1990年 | 5篇 |
1988年 | 6篇 |
1986年 | 3篇 |
1985年 | 10篇 |
1984年 | 18篇 |
1983年 | 15篇 |
1982年 | 15篇 |
1981年 | 13篇 |
1980年 | 4篇 |
1979年 | 8篇 |
1978年 | 7篇 |
1977年 | 10篇 |
1976年 | 14篇 |
1975年 | 10篇 |
1974年 | 3篇 |
1973年 | 4篇 |
1972年 | 2篇 |
排序方式: 共有1478条查询结果,搜索用时 15 毫秒
1.
《Process Biochemistry》2014,49(5):882-889
The VP4 protein of infectious bursal disease virus (IBDV) is a serine protease that processes the polyprotein for viral assembly. VP4 has been found to associate primarily with type II IBDV tubules that are 24 nm in diameter. In this study, a chimeric VP4, assigned as HS1VP4, was constructed with a VP4-autocleavage site inserted between the N-terminal His-tag and the VP4 sequence. The results showed that the VP4 forms tubules after the self-cleavage of HS1VP4 when expressed in Escherichia coli. Furthermore, a deletion of 28 amino acids at the C-terminus of VP4 resulted in monomers and dimers instead of tubule formation; mutants of S652A and K692A at active site destroyed the activity. The endopeptidase activity of these monomers and dimers was approximately 12.5 times higher than that of VP4 tubules. Additionally, the formation of tubules inhibited VP4 protease activity, as demonstrated through in vitro assays. The production and characterization of monomers or dimers that have greater endopeptidase activity and protease activity than tubules can provide further insight into VP4 tubule assembly and the regulation of VP4 activity in host cells; this insight will facilitate the development of new anti-IBDV strategies. 相似文献
2.
Laboratory incubation and field experiments were conducted to evaluate thiourea, ATC (4-amino-1, 2, 4 triazole hydrochloride)
and N-Serve 24 E (2-chloro-6-trichloromethyl-pyridine) as inhibitors of nitrification of fertilizer N. In the incubation experiment,
most of the added aqueous NH3 or urea was nitrified at 14 days on both soils, but addition of the inhibitors to fertilizer N decreased the conversion of
NH4−N to NO3−N markedly. There was less nitrification for ATC and thiourea but not for N-Serve 24 E when the fertilizers and the inhibitors
were placed at a point as opposed to when mixed into soil. After 28 days, ATC and N-Serve 24 E were more effective in inhibiting
nitrification than thiourea. ATC and N-Serve 24 E also inhibited release of mineral N (NH4−N+NO3−N) from native soil N. In the uncropped field experiment, which received N fertilizers in the fall, nitrification of fall-applied
N placed in the 15-cm bands was almost complete by early May in the Malmo soil, but not in the Breton soil. When ATC or thiourea
had been applied with urea, nitrification of fall-applied N was depressed by May and the recovery of applied N as NH4−N was greater with increasing band spacing to 60 cm or placing N fertilizer in nests (a method of application where urea
prills were placed at a point in the soil in the center of 60×60 cm area). In late June, the percentage recovery of fall-applied
N in soil as NH4−N or mineral N increased with wide band spacing, or nest placement, or by adding ATC to fertilizer N on both soils. These
results indicate that placing ammonium-based N fertilizers in widely-spaced bands or in nests with low rates of inhibitors
slows nitrification enough to prevent much of the losses from fall-applied N.
Scientific Paper No. 552, Lacombe Research Station, Research Branch, Agric, Can. 相似文献
3.
4.
Introduction of T-dependent antigens into the subarachnoid space (isas) resulted in higher systemic antibody responses in mice than injections into the peritoneal cavity (ip) or other sites commonly used for immunization. Antibody production in isas immunized mice was not increased by treatment with cyclophosphamide (Cy) at doses known to abolish T-suppressor-cell activity, but such treatment increased antibody production in ip immunized mice toward the higher level which was observed in the isas immunized animals. Suppressor cell-dependent low zone tolerance (LZT) to TNP-BSA could not be induced by isas injections of deaggregated BSA (d-BSA). Conversely, mice which were unresponsive to ip injected d-BSA showed consistent systemic antibody responses when the antigen was injected isas. These observations indicate that immune responses initiated within the CNS are associated with relatively ineffective induction of systemic suppressor cell activity. 相似文献
5.
Uncertainties surrounding the structures of the Δ7-sterols in the seeds of Cucurbita maxima have been resolved. Seven components were found by TLC, GLC, HPLC, mass spectrometry and 1H NMR. They were 24β-ethyl-5α-cholesta-7,22,25(27)-trien-3β-ol, 24β-ethyl-5α-cholesta-7,25(27)-dien-3gb-ol, avenasterol, spinasterol, 24-dihydrospinasterol, 24ζ-methyllathosterol and 25(27)-dehydrofungisterol. The 1H NMR spectra indicated that the sterols with an ethyl substituent at C-24 occurred in the absence of their C-24 epimers. This seems to be the first instance of the detection of 25(27)-dehydrofungisterol in a higher plant. 相似文献
6.
The irreversible binding of [2,3-14C]acrylonitrile (VCN) to proteins, RNA and DNA of various tissues of male Sprague-Dawley rats after a single oral dose of 46.5 mg/kg (0.5 LD50) has been studied. Proteins were isolated by chloroform-isoamyl alcohol-phenol extraction. RNA and DNA were separated by hydroxyapatite chromatography. Binding of VCN to proteins was extensive and was time dependent. Radioactivity in nucleic acids was registered in the liver and the target organs, stomach and brain. DNA alkylation, which increased by time, was significantly higher in the target organs, brain and stomach (119 and 81 pmol/mg, respectively, at 24 h) than that in the liver. The covalent binding indices for the liver, stomach and brain at 24 h after dosing were, 5.9, 51.9 and 65.3, respectively. These results suggest that VCN is able to act as a multipotent carcinogen by alkylation of DNA in the extrahepatic target tissues, stomach and brain. 相似文献
7.
Immunologic and clinical improvement of progressive coccidioidomycosis following administration of transfer factor 总被引:8,自引:0,他引:8
Three patients with progressive coccidioidomycosis were given preparations of transfer factor (TF). Adverse reactions to TF were minimal. Following TF administration two of these patients had prolonged clinical remissions in their coccidioidal disease. Cellular immune responses were sequentially evaluated by coccidioidininduced delayed-type skin tests, lymphocyte blast transformation and macrophage inhibition factor production (MIF). These three patients each exhibited different cellular immune patterns before and after TF administration. Two patients converted their coccidioidin skin tests, and one converted lymphocyte transformation response to coccidioidin. Also, TF apparently favorably affected the MIF response in all three patients. 相似文献
8.
Kerry R. Emslie M. Barrie Coukell Debra Birch Keith L. Williams 《Journal of biotechnology》1996,50(2-3):149-159
We have previously reported expression of the rotavirus outer capsid glycoprotein, VP7, in the relatively new expression host, Dictyostelium discoideum. To optimise yields of recombinant VP7, we examined the role of Ca2+ since stability of both VP7 and mature rotavirus during a rotavirus infection are calcium-dependent. Low micromolar levels of free extracellular Ca2+ were required to maximise yields of VP7 in D. discoideum whilst levels of VP7 were reduced following depletion of intracellular Ca2+ reserves using A23187 and EGTA. Immunoblot analysis suggested that VP7 was being degraded in an intracellular compartment. Immunoprecipitation with a conformation-dependent neutralising antibody confirmed that EGTA-induced Ca2+ chelation alters the conformation of VP7. These results suggest that stability of VP7 is dependent on maintaining adequate levels of intracellular Ca2+ and that conformational changes in VP7 which occur following depletion of Ca2+ reserves induce rapid proteolysis of the protein. Since these results establish conditions for expressing optimal levels of VP7 in the correct conformation they have important implications for the development of a subunit vaccine based on recombinant VP7. 相似文献
9.
PreparationofMeioticKarytypeofMouseOocyteLiChaojunYanLeipingZhangXiranChenYifeng(BiologyDepartmentofNanjingNormalUniversity,Nanjing210097)哺乳动物的卵母细胞的减数分裂过程中存在两次自发的停滞现象,第一次是在第一次减数分裂前期的双线期,这一静止期持续很长时间,一直到动物性成熟后卵母细胞进入发有周刎,在保住腺激素的作用下,卵母细胞的第一次减数分裂才重新启动.完成第一次减数分裂后.又停滞在第二次减数分裂的中期,在椅子或化学因素刺激的作用下,完成第二次减数分裂[4].因此,对哺乳动物的卵母细胞在一… 相似文献
10.
A组轮状病毒SA11VP6基因的克隆和表达 总被引:4,自引:0,他引:4
从SA11VP6基因全序列克隆开始,设计一对两端带有酶切位点的引物,逆转录PCR扩增出VP6全基因CDNA。经酶切后插入PUC19,构建了VP6全基因克隆PRA6。再经酶切后插入痘苗病毒载休质凿PJSA1175中。利用Lipofectin导入TK143细胞,利用TK基因和Lac基因作为重组病毒的筛选标记。表达产物用单克隆抗体ELISA法检测,发现细胞培养上清和细胞裂解液都是阳性。Western b 相似文献