全文获取类型
收费全文 | 4210篇 |
免费 | 163篇 |
国内免费 | 218篇 |
专业分类
4591篇 |
出版年
2023年 | 47篇 |
2022年 | 71篇 |
2021年 | 77篇 |
2020年 | 71篇 |
2019年 | 104篇 |
2018年 | 112篇 |
2017年 | 71篇 |
2016年 | 82篇 |
2015年 | 84篇 |
2014年 | 239篇 |
2013年 | 306篇 |
2012年 | 198篇 |
2011年 | 277篇 |
2010年 | 182篇 |
2009年 | 194篇 |
2008年 | 242篇 |
2007年 | 256篇 |
2006年 | 183篇 |
2005年 | 166篇 |
2004年 | 109篇 |
2003年 | 108篇 |
2002年 | 93篇 |
2001年 | 66篇 |
2000年 | 60篇 |
1999年 | 93篇 |
1998年 | 58篇 |
1997年 | 53篇 |
1996年 | 49篇 |
1995年 | 56篇 |
1994年 | 45篇 |
1993年 | 46篇 |
1992年 | 56篇 |
1991年 | 57篇 |
1990年 | 42篇 |
1989年 | 34篇 |
1988年 | 29篇 |
1987年 | 47篇 |
1986年 | 18篇 |
1985年 | 60篇 |
1984年 | 57篇 |
1983年 | 52篇 |
1982年 | 42篇 |
1981年 | 41篇 |
1980年 | 50篇 |
1979年 | 34篇 |
1978年 | 35篇 |
1977年 | 24篇 |
1976年 | 29篇 |
1975年 | 27篇 |
1973年 | 22篇 |
排序方式: 共有4591条查询结果,搜索用时 15 毫秒
101.
102.
Optimization of sulfated modification conditions of tremella polysaccharide and effects of modifiers on cellular infectivity of NDV 总被引:2,自引:0,他引:2
Zhao X Hu Y Wang D Guo L Yang S Fan Y Zhao B Wang Y Abula S 《International journal of biological macromolecules》2011,49(1):44-49
Based on our previous research, sulfated modification conditions of Tremella polysaccharide (TPS), the chlorosulfonic acid to pyridine (CSA-Pry) ratio, reaction temperature and time, were optimized by L9 (34) orthogonal design taking the yield and degree of sulfation (DS) of modifiers as indexes. Two TPSs, TPStp and TPS70c, were modified under optimized conditions. The effects of two modifiers, sTPStp and sTPS70c, on cellular infectivity of NDV were determined by MTT method taking the non-modified TPStp, TPStc and TPS70c as controls. The results showed that the optimized modification conditions were reaction temperature of 80 °C, CSA-Pry ratio of 1:6 and reaction time of 1.5 h. Five polysaccharides at proper concentrations could significantly inhibit the infectivity of NDV to CEF. The virus inhibitory rates of sTPStp at 1.563 μg mL−1 group were the highest and significantly higher than those of other three non-modified polysaccharide groups in three sample-adding modes. This indicated that sulfated modification could significantly improve the antiviral activity of TPS. sTPStp possessed the best efficacy and would be as a component of antiviral polysaccharide drug. 相似文献
103.
Importance of rare taxa for bacterial diversity in the rhizosphere of Bt- and conventional maize varieties 总被引:1,自引:0,他引:1
Anja B Dohrmann Meike Küting Sebastian Jünemann Sebastian Jaenicke Andreas Schlüter Christoph C Tebbe 《The ISME journal》2013,7(1):37-49
Ribosomal 16S rRNA gene pyrosequencing was used to explore whether the genetically modified (GM) Bt-maize hybrid MON 89034 × MON 88017, expressing three insecticidal recombinant Cry proteins of Bacillus thuringiensis, would alter the rhizosphere bacterial community. Fine roots of field cultivated Bt-maize and three conventional maize varieties were analyzed together with coarse roots of the Bt-maize. A total of 547 000 sequences were obtained. Library coverage was 100% at the phylum and 99.8% at the genus rank. Although cluster analyses based on relative abundances indicated no differences at higher taxonomic ranks, genera abundances pointed to variety specific differences. Genera-based clustering depended solely on the 49 most dominant genera while the remaining 461 rare genera followed a different selection. A total of 91 genera responded significantly to the different root environments. As a benefit of pyrosequencing, 79 responsive genera were identified that might have been overlooked with conventional cloning sequencing approaches owing to their rareness. There was no indication of bacterial alterations in the rhizosphere of the Bt-maize beyond differences found between conventional varieties. B. thuringiensis-like phylotypes were present at low abundance (0.1% of Bacteria) suggesting possible occurrence of natural Cry proteins in the rhizospheres. Although some genera indicated potential phytopathogenic bacteria in the rhizosphere, their abundances were not significantly different between conventional varieties and Bt-maize. With an unprecedented sensitivity this study indicates that the rhizosphere bacterial community of a GM maize did not respond abnormally to the presence of three insecticidal proteins in the root tissue. 相似文献
104.
【背景】碱性丝氨酸蛋白酶(Subtilisin)是一种具有广泛用途的工业酶制剂。【目的】旨在通过优化启动子、信号肽及培养基组分来提高地衣芽胞杆菌中碱性丝氨酸蛋白酶产量。【方法】以地衣芽胞杆菌BL10为出发菌株,构建了含有4种不同类型启动子(PbacA、P43、PaprE和PsrfA)及4种不同类型信号肽(SPVpr、SPSacB、SPSacC和SPAprE)的碱性丝氨酸蛋白酶表达菌株,并在获得高产菌株的基础上进行培养基优化。【结果】4种启动子的表达水平为PbacAPaprEP43PsrfA,4种信号肽的分泌效率为SPAprESPSacCSPSacBSPVpr。其中,菌株BL10/pPbacA-aprE产生最高的碱性丝氨酸蛋白酶酶活(275.21 U/mL),相比于出发菌株BL10/pHY-aprE (167.98 U/mL)提高了64%。随后,通过对发酵培养基成分进行优化并结合正交优化,获得了一种高产碱性丝氨酸蛋白酶的培养基(g/L):玉米淀粉40.0,豆粕50.0,(NH4)2SO4 4.0,K2HPO4 3.0,CaCO3 1.0。最后,碱性丝氨酸蛋白酶酶活提高到747.37 U/mL,是初始酶活的4.45倍。【结论】为工业化高产碱性丝氨酸蛋白酶提供了一种有效策略。 相似文献
105.
【背景】猪水肿病大肠杆菌引发的疾病造成了很大的危害,但现有培养基存在培养密度低的问题。【目的】研制出高抗原活性猪水肿病大肠杆菌疫苗培养基。【方法】以常用的市售猪水肿培养基为对照,通过单因素试验、爬坡试验(Plackett-Burman, PB)、响应面(Box-Behnken, BB)试验对猪水肿培养基进行响应面优化,得到猪水肿培养基最优配方。以响应面试验得到的培养基培养猪水肿病大肠杆菌,评价不同培养时间点菌株的抗原活性,制作灭活疫苗,进行动物免疫保护试验。【结果】对研制的培养基进行扩大培养验证,发现扩大培养得到的菌株活菌数可达5×109 CFU/mL以上,约为对照组的2倍。制备的灭活疫苗效价可达1:140 000,并在9 h时抗原蛋白效价达到最高。【结论】本研究研制出的疫苗培养基显著提高了猪大肠杆菌菌体密度,并可提高菌体抗原活性,为猪水肿病灭活疫苗的制备提供了技术指引。 相似文献
106.
【目的】白酒可培养微生物的分离筛选是白酒行业重要的研究内容,本文旨在构建中国白酒生产环境中可培养微生物信息数据库。【方法】数据库信息主要来源于通过人工查阅和整理目前已发表的白酒微生物的相关的文献报道和菌种保藏中心相关的筛选自白酒生态系统的微生物信息。数据库主要设计3个功能:(1)白酒可培养微生物信息检索:通过菌株名称、分离位置、培养基、代谢产物等为条件检索相关的白酒可培养微生物信息,从而获取该白酒微生物详细的生理生化与分类学信息;(2)培养基信息检索:通过特定培养基成分,培养基编号和名称检索相关的培养基信息,包括培养的组成和配制方法。(3)数据更新:在线上传新的可培养微生物和培养基信息。【结果】目前数据库共收1221种白酒可培养微生物和295种培养基信息,网址为:http://cmbaijiu.i-sanger.com/。【结论】本数据库是我国白酒领域首个可培养微生物信息的数据库,将有助于白酒微生物培养的相关研究工作开展。 相似文献
107.
108.
109.
Reptilian transferrins: evolution of disulphide bridges and conservation of iron-binding center 总被引:1,自引:0,他引:1
Transferrins, found in invertebrates and vertebrates, form a physiologically important family of proteins playing a major role in iron acquisition and transport, defense against microbial pathogens, growth and differentiation. These proteins are bilobal in structure and each lobe is composed of two domains divided by a cleft harboring an iron atom. Vertebrate transferrins comprise of serotransferrins, lactoferrins and ovotransferrins. In mammals serotransferrins transport iron in physiological fluids and deliver it to cells, while lactoferrins scavenge iron, limiting its availability to invading microbes. In oviparous vertebrates there is only one transferrin gene, expressed either in the liver to be delivered to physiological fluids as serotransferrin, or in the oviduct with a final localization in egg white as ovotransferrin. Being products of one gene sero- and ovotransferrin are identical at the amino-acid sequence level but with different, cell specific glycosylation patterns. Our knowledge of the mechanisms of transferrin iron binding and release is based on sequence and structural data obtained for human serotransferrin and hen and duck ovotransferrins. No sequence information about other ovotransferrins was available until our recent publication of turkey, ostrich, and red-eared turtle (TtrF) ovotransferrin mRNA sequences [Ciuraszkiewicz, J., Olczak, M., Watorek, W., 2006. Isolation, cloning and sequencing of transferrins from red-eared turtle, African ostrich and turkey. Comp. Biochem. Physiol. 143 B, 301-310]. In the present paper, ten new reptilian mRNA transferrin sequences obtained from the Nile crocodile (NtrF), bearded dragon (BtrF), Cuban brown anole (AtrF), veiled and Mediterranean chameleons (VtrF and KtrF), sand lizard (StrF), leopard gecko (LtrF), Burmese python (PtrF), African house snake (HtrF), and grass snake (GtrF) are presented and analyzed. Nile crocodile and red-eared turtle transferrins have a disulphide bridge pattern identical to known bird homologues. A partially different disulphide bridge pattern was found in the Squamata (snakes and lizards). The possibility of a unique interdomain disulphide bridge was predicted for LtrF. Differences were found in iron-binding centers from those of previously known transferrins. Substitutions were found in the iron-chelating residues of StrF and TtrF and in the synergistic anion-binding residues of NtrF. In snakes, the transferrin (PtrF, HtrF and GtrF) N-lobe "dilysine trigger" occurring in all other known transferrins was not found, which indicates a different mechanism of iron release. 相似文献
110.
Koichiro Yoshihara Motokatsu Tsuyuki Asako Itaya Yasuharu Tanaka Tomoya Kamiya 《Molecular and cellular biochemistry》1994,135(2):143-151
HL-60 cells transferred from serum-supplemented to serum-free culture medium initially bound to culture plate tightly and then released from the plate on increasing the culture time and resumed exponential growth after about 8 h lag. At the initial stage of the culture, the cells became extremely sensitive to 3-aminobenzamide, a potent inhibitor of poly (ADP-ribose) polymerase, and, at 1 mM, 80 to 90% of the cells were lysed within 20 h, whereas the inhibitor was totally ineffective on the cell growth in serum-supplemented medium at the concentration. Non-inhibitory analogs of the inhibitor were ineffective. Assay of poly(ADP-ribose) polymerase activity in permeable cells indicated that a transient activation of the enzyme occurred during the culture in serum-free medium (the maximum activation was observed at 8 h of the culture). The cells conditioned in serum-free medium for 24 h acquired significant resistancy to the inhibitor. A low concentration of fibronectin (5 to 10/ml) and a relatively high concentration of bovine serum albumin (0.5 to 1 mg/ml) effectively blocked the cell attachment to plate and also the 3-aminobenzamide-induced cell lysis. These results suggest that poly(ADP-ribose) polymerase is involved in a process essential for HL-60 cells to adapt to a serumdeprived growth condition. 相似文献