全文获取类型
收费全文 | 4881篇 |
免费 | 193篇 |
国内免费 | 369篇 |
出版年
2023年 | 34篇 |
2022年 | 60篇 |
2021年 | 65篇 |
2020年 | 56篇 |
2019年 | 109篇 |
2018年 | 107篇 |
2017年 | 77篇 |
2016年 | 85篇 |
2015年 | 135篇 |
2014年 | 332篇 |
2013年 | 319篇 |
2012年 | 298篇 |
2011年 | 359篇 |
2010年 | 268篇 |
2009年 | 281篇 |
2008年 | 291篇 |
2007年 | 323篇 |
2006年 | 286篇 |
2005年 | 241篇 |
2004年 | 200篇 |
2003年 | 179篇 |
2002年 | 111篇 |
2001年 | 104篇 |
2000年 | 105篇 |
1999年 | 117篇 |
1998年 | 91篇 |
1997年 | 86篇 |
1996年 | 75篇 |
1995年 | 88篇 |
1994年 | 78篇 |
1993年 | 62篇 |
1992年 | 57篇 |
1991年 | 58篇 |
1990年 | 55篇 |
1989年 | 31篇 |
1988年 | 30篇 |
1987年 | 19篇 |
1986年 | 27篇 |
1985年 | 15篇 |
1984年 | 12篇 |
1983年 | 10篇 |
1982年 | 10篇 |
1981年 | 12篇 |
1980年 | 11篇 |
1979年 | 7篇 |
1978年 | 10篇 |
1977年 | 8篇 |
1976年 | 13篇 |
1972年 | 7篇 |
1971年 | 6篇 |
排序方式: 共有5443条查询结果,搜索用时 15 毫秒
991.
鲤鱼肌肉生长抑制素基因(MSTN)的克隆及其组织表达特征 总被引:5,自引:0,他引:5
肌肉生长抑制素(Myostatin,MSTN)是动物肌肉发育和生长过程中的负调控因子,对MSTN的研究将有助于促进动物生产。鲤鱼是我国的主要淡水养殖对象之一。因此,我们采用RT-PCR方法克隆了鲤鱼MSTN cDNA(No.EF551058)的部分序列,长度为921bp,编码306个氨基酸残基。鲤鱼MSTN具有MSTN的共同特征,有蛋白酶水解位点RIRR和9个保守的半胱氨酸残基。多重序列比较发现其与斑马鱼GDF8有极近的亲缘关系,96.7%的氨基酸序列同源。不同组织的RT-PCR分析发现鲤鱼MSTN主要在肌肉和脑部表达,而其他所检测组织未见表达。鲤鱼MSTN不仅在肌肉生长发育中发挥作用,可能在神经系统发育中也有其作用。 相似文献
992.
将编码融合蛋白GST-SUMO-MT的DNA片段连接到大肠杆菌表达载体pET-28a中,构建重组表达质粒pET-GS-MT并转化到大肠杆菌Origami(DE3)中。20℃,1mM的IPTG诱导20h后,获得分子量约为43Kd的融合蛋白,表达量占菌体上清总蛋白的38.4%。利用谷胱甘肽交联琼脂糖(Glutathione Sepharose 4B)凝胶柱和Sephardex G-25分子筛联用可以得到纯度为95%以上的融合蛋白,得率约为70mg/L。该融合蛋白可与GST抗体产生阳性反应。融合蛋白GST-SUMO-MT可以显著提高宿主对Cd2+、Zn2+和Cu2+离子聚积的能力,其耐受能力比对照组分别提高4.2倍、4倍及1.6倍。此外,原子吸收光谱法测定,每分子GST-SUMO-MT可以结合2-3个Cd2+离子。 相似文献
993.
To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae. 相似文献
994.
两种药物性肝炎诊断评分标准的临床应用分析 总被引:1,自引:0,他引:1
目的:比较Maria药物性肝损害(1997年)诊断标准和我国常用诊断标准对药物性肝损害的效价。方法:回顾性分析我院2005年1月至2006年4月期间经肝穿病理诊断为药物性肝损害的54例病例,分别按照上述两个标准评分。评价每一标准的准确性。结果:符合Maria药物性肝损害(1997年)诊断标准的(>10分)有45例,占83%;符合我国常用诊断标准的有39例,占72%。结论:两个诊断标准符合率较高,均可做为药物性肝损害的诊断标准,但仍有部分病例不能准确诊断,需进一步完善。 相似文献
995.
目的:获得肠三叶因子(ITF)的原核表达产物及抗rITF抗体,为深入研究ITF的作用机制及其受体研究奠定基础。方法:常规提取人小肠组织总RNA,用RT-PCR获得ITF编码基因片段,克隆至质粒pET32a获得原核表达栽体,双酶切和测序后转化至Origami B(DE3)用IPTG诱导表达,优化条件获得最大表达产量;用SDS-PAGE、Western blot鉴定表达产物,亲和层析纯化获得的重组蛋白rITF皮下多点注射家兔,制备多克隆抗体,并用此抗体进行大鼠肠组织免疫组化研究。结果:测序证实PCR扩增获得ITF全长基因序列与基因文库中的完全一致,将该基因片段正确插入表达载体pET32a中、优化表达条件后,重组蛋白的表达量达到50mg/L;Western blot证明重组蛋白具有良好的抗原性和特异性;通过Ni-NTA亲和层析、超滤离心后,得到90%纯度的蛋白;收集兔血清,纯化后获得特异性良好的ITF抗体,免疫组化染色肠组织显示ITF表达的部位定位于杯状细胞。结论:成功构建了表达载体pET32a-ITF,在大肠杆菌中表达并纯化获得纯度较高的rITF,并获得了生物活性较高的ITF抗体,ITF主要在肠道杯状细胞分泌表达。 相似文献
996.
997.
998.
Various approaches have been developed to define conservation units for plant and animal species. In this study we combined
nuclear microsatellites (from a previous published study) and chloroplast microsatellites (assessed in the present study),
leaf and seed morphology traits and abiotic variables (climate and soil) to define evolutionary significant units (ESU) of
Santalum austrocaledonicum, a tree species growing in New Caledonia. Results for chloroplast microsatellites showed that the total population heterozygosity
was␣high, (H
cp = 0.84) but varied between islands. Differentiation was strong in the total population (F
stcp = 0.66) but also within the main island Grande Terre (F
stcp = 0.73) and within Iles Loyauté (F
stcp = 0.52), highlighting a limited gene flow between populations. These results confirmed those obtained with nuclear microsatellites.
The cluster analysis on molecular markers discriminated two main groups constituted by the populations of Grande Terre and
the populations of Iles Loyauté. A principal component analysis of leaf and seed morphology traits singled out the populations
of Iles Loyauté and the western populations of Grande Terre. Quantitative genetic analyses showed that the variation between
populations was under genetic control (broad sense heritability close to 80%). A high correlation between rainfall and morphological
traits suggested an impact of climate on this variation. The integration of these results allows to define two ESUs, one corresponding
to Grande Terre and Ile des Pins and the other the Iles Loyauté archipelago. This study stresses the need to restore some
populations of Grande Terre that are currently threatened by their small size. 相似文献
999.
1000.
Wen-rong YAO Guo-qiang PAN Cheng-long XIONG Qian-fu ZHOU Qi-you XIAO Ming-hui LI Yong-zhen ZHANG 《Virologica Sinica》2007,22(4):307-315
Saliva and blood were collected from two patients who had not received post exposure prophylaxis in the cities of Wenzhou and Xinning respectively. Both patients were confirmed as positive for rabies by detection of rabies virus specific nucleoprotein antibodies in the sera by Western Blot. However, rabies virus specific RNA was only identified in the saliva collected from the patient in Wenzhou. Furthermore, the isolate Zhejiang Wz0 (H) was obtained by inoculating one-day-old suckling mice. Both nucleoprotein (N) and glycoprotein (G) genes from the isolate were amplified by RT-PCR and sequenced. Phylogenetic analysis indicated that the isolate belonged to classic rabies virus, and shared a higher homology with the street viruses from dogs in the main endemic areas in China and the street virus from dogs in Indonesia than with other known strains. Further comparison of the deduced amino acid sequences between the isolate and the vaccine strains used in China showed that the virus had a higher level of homology with the vaccine strain CTN than with the other vaccine strains (3aG, PV, PM and ERA). In particular, amino acid residues substitutions located in antigenic site Ⅲ in the G protein, which could react with the neutralizing antibodies, were observed. These results suggested that the virus belonged to the classic rabies virus, and both N and G genes diverged from the current vaccine strains used in China at either the nucleotide or the amino acid level. 相似文献