全文获取类型
收费全文 | 15027篇 |
免费 | 1367篇 |
国内免费 | 1649篇 |
专业分类
18043篇 |
出版年
2024年 | 62篇 |
2023年 | 256篇 |
2022年 | 557篇 |
2021年 | 907篇 |
2020年 | 642篇 |
2019年 | 746篇 |
2018年 | 711篇 |
2017年 | 508篇 |
2016年 | 680篇 |
2015年 | 996篇 |
2014年 | 1127篇 |
2013年 | 1252篇 |
2012年 | 1386篇 |
2011年 | 1300篇 |
2010年 | 767篇 |
2009年 | 717篇 |
2008年 | 779篇 |
2007年 | 663篇 |
2006年 | 655篇 |
2005年 | 427篇 |
2004年 | 417篇 |
2003年 | 391篇 |
2002年 | 330篇 |
2001年 | 241篇 |
2000年 | 196篇 |
1999年 | 197篇 |
1998年 | 164篇 |
1997年 | 117篇 |
1996年 | 110篇 |
1995年 | 97篇 |
1994年 | 76篇 |
1993年 | 65篇 |
1992年 | 86篇 |
1991年 | 58篇 |
1990年 | 52篇 |
1989年 | 52篇 |
1988年 | 43篇 |
1987年 | 28篇 |
1986年 | 22篇 |
1985年 | 28篇 |
1984年 | 19篇 |
1983年 | 11篇 |
1982年 | 17篇 |
1981年 | 11篇 |
1980年 | 8篇 |
1979年 | 11篇 |
1978年 | 10篇 |
1977年 | 7篇 |
1975年 | 11篇 |
1973年 | 7篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
81.
鸡生长激素基因5′端部分调控区的克隆分析 总被引:4,自引:0,他引:4
利用PCR技术扩增、克隆、测序了7个跨越不同生长速度的鸡品种(系)的生长激素(GH)基因的5′端部分调控区。这7个品种(系)分别为:高生长速度的宝罗肉鸡父本;较高生长速度和一定的产蛋性能的宝罗肉鸡母本;肉蛋兼用型的芦花鸡;中等生长速度和中等体重的蛋鸡品种洛岛红和农大褐;生长速度较慢体重较轻的蛋鸡品种北京白鸡和生长速度很慢但又不是矮小型的地方品种丝毛乌骨鸡。所扩增的片段长度为760bp,包括了转录起始位点5′端侧翼区的473bp、两个可能的TATA框、组织特异性转录因子结合位点、第一个外显子和部分第一内含子。所有克隆的鸡GH基因5′端调控区与Tanaka发表的序列相比,均在-34碱基的位置上缺失一个胞嘧啶C,在 160与 161碱基之间多1个胸腺嘧啶丁,在 175碱基的位置上出现了以腺嘌呤A替换了鸟嘌呤G的情况。7个品种(系)之间的比较显示,鸡GH基因启动区具有极高的保守性,在具有不同生长速度的鸡品种间不存在任何差异。说明鸡的不同生长速度和成年体重,不是由于生长激素5′调控区变异造成的。鸡生长激素基因表达的差异可能受到内含子或3′端侧翼序列的影响。 相似文献
82.
A marine fish cell line from the snout of red spotted grouper Epinephelus akaara, a protogynous hermaphrodite, was established, characterized, and subcultured with more than 60 passages. The grouper snout
cell line (GSC) cells multiplied well in Dulbecco’s modified Eagle’s medium (DMEM) medium supplemented with 10% fetal bovine
serum. The optimal growth temperature was 25°C, and morphologically the cells were fibroblastic. Chromosome analysis revealed
that the GSC cell line has a normal diploid karyotype with
. A virus titration study indicated that the cells were susceptible to turbot Scophthalmus Maximus rhabdovirus (SMRV) (108.5 TCID50 ml−1), while the viral titer of frog Rana grylio virus 9807 (RGV9807) reached 103.5 TCID50 ml−1. The infection was confirmed by cytopathic effect (CPE), immunofluorescence, and electron microscopy experiments, which detected
the viral particles in the cytoplasm of virus-infected cells, respectively. Further, significant fluorescent signals were
observed when the GSC cells were transfected with pEGFP vector DNA, indicating their potential utility for transgenic and
genetic manipulation studies. 相似文献
83.
84.
Ying Wang Yunjing Zhang Xinwan Su Qiongzi Qiu Yuan Yuan Chunhua Weng Sailan Zou Yan Tian Weidong Han Pengyuan Liu Xingyi Guo Jianhua Mao Xianghui Fu Ping Wang Weiqiang Lin 《International journal of biological sciences》2022,18(4):1491
Clear cell renal cell carcinoma (ccRCC) is a primary kidney cancer with high aggressive phenotype and extremely poor prognosis. Accumulating evidence suggests that circular RNAs (circRNAs) play pivotal roles in the occurrence and development of various human cancers. However, the expression, clinical significance and regulatory role of circRNAs in ccRCC remain largely unclear. Here we report that circDVL1 to be reduced in the serums and tissues from ccRCC patients, and to negatively correlate with ccRCC malignant features. Overexpression of circDVL1 inhibits proliferation, induces G1/S arrest, triggers apoptosis, and reduces migration and invasion in different ccRCC cells in vitro. Correspondingly, circDVL1 overexpression suppresses ccRCC tumorigenicity in a mouse xenograft model. Mechanistically, circDVL1 serves as a sponge for oncogenic miR-412-3p, thereby preventing miR-412-3p-mediated repression of its target protocadherin 7 (PCDH7) in ccRCC cells. Collectively, our results demonstrate that circDVL1 exerts tumor-suppressive function during ccRCC progression through circDVL1/miR-412-3p/PCDH7 axis, and suggest that circDVL1 could be a novel diagnostic and prognositc marker and therapeutic target for ccRCC. 相似文献
85.
Lei Yang Lingqian Tian Leshan Li Qiuhong Liu Xiang Guo Yuan Zhou Rongjuan Pei Xinwen Chen Yun Wang 《中国病毒学》2022,37(3):341-347
Transformation-associated recombination (TAR) has been widely used to assemble large DNA constructs. One of the significant obstacles hindering assembly efficiency is the presence of error-prone DNA repair pathways in yeast, which results in vector backbone recircularization or illegitimate recombination products. To increase TAR assembly efficiency, we prepared a dual-selective TAR vector, pGFCS, by adding a PADH1-URA3 cassette to a previously described yeast-bacteria shuttle vector, pGF, harboring a PHIS3-HIS3 cassette as a positive selection marker. This new cassette works as a negative selection marker to ensure that yeast harboring a recircularized vector cannot propagate in the presence of 5-fluoroorotic acid. To prevent pGFCS bearing ura3 from recombining with endogenous ura3-52 in the yeast genome, a highly transformable Saccharomyces cerevisiae strain, VL6-48B, was prepared by chromosomal substitution of ura3-52 with a transgene conferring resistance to blasticidin. A 55-kb genomic fragment of monkeypox virus encompassing primary detection targets for quantitative PCR was assembled by TAR using pGFCS in VL6-48B. The pGFCS-mediated TAR assembly showed a zero rate of vector recircularization and an average correct assembly yield of 79% indicating that the dual-selection strategy provides an efficient approach to optimizing TAR assembly. 相似文献
86.
Hang Yang Huijun Yuan Xiaohui Zhao Meng Xun Shangrui Guo Nan Wang Bing Liu Hongliang Wang 《中国病毒学》2022,37(3):380-389
The recent COVID-19 pandemic poses a global health emergency. Cellular entry of the causative agent SARS-CoV-2 is mediated by its spike protein interacting with cellular receptor-human angiotensin converting enzyme 2 (ACE2). Here, by using lentivirus based pseudotypes bearing spike protein, we demonstrated that entry of SARS-CoV-2 into host cells was dependent on clathrin-mediated endocytosis, and phosphoinositides played essential roles during this process. In addition, we showed that the intracellular domain and the catalytic activity of ACE2 were not required for efficient virus entry. Finally, we showed that the current predominant Delta variant, although with high infectivity and high syncytium formation, also entered cells through clathrin-mediated endocytosis. These results provide new insights into SARS-CoV-2 cellular entry and present proof of principle that targeting viral entry could be an effective way to treat different variant infections. 相似文献
87.
Dan Chen Satoko Ito Hong Yuan Toshinori Hyodo Kenji Kadomatsu Michinari Hamaguchi Takeshi Senga 《Cell cycle (Georgetown, Tex.)》2015,14(10):1529-1539
Echinoderm microtubule-associated protein (EMAP)-like (EML) family proteins are microtubule-associated proteins that have a conserved hydrophobic EMAP-like protein (HELP) domain and multiple WD40 domains. In this study, we examined the role of EML4, which is a member of the EML family, in cell division. Time-lapse microscopy analysis demonstrated that EML4 depletion induced chromosome misalignment during metaphase and delayed anaphase initiation. Further analysis by immunofluorescence showed that EML4 was required for the organization of the mitotic spindle and for the proper attachment of kinetochores to microtubules. We searched for EML4-associating proteins by mass spectrometry analysis and found that the nuclear distribution gene C (NUDC) protein, which is a critical factor for the progression of mitosis, was associated with EML4. This interaction was mediated by the WD40 repeat of EML4 and by the C-terminus of NUDC. In the absence of EML4, NUDC was no longer able to localize to the mitotic spindle, whereas NUDC was dispensable for EML4 localization. Our results show that EML4 is critical for the loading of NUDC onto the mitotic spindle for mitotic progression. 相似文献
88.
Miao Liu Yanfei Ru Yihua Gu Jianan Tang Tiancheng Zhang Jun Wu Fudong Yu Yao Yuan Chen Xu Jian Wang Huijuan Shi 《Biochimica et Biophysica Acta (BBA)/General Subjects》2018,1862(3):660-668
Background
We previously cloned the Ssp411 gene. We found that the Ssp411 protein is predominantly expressed in elongated spermatids in the rat testis in a stage-dependent manner. Although our findings strongly suggested that Ssp411 might play an important role in mammalian spermatogenesis, this hypothesis has not been studied.Methods
We first used real-time PCR, Western blotting and immunohistochemistry to confirm that the expression pattern of Ssp411 in several murine tissues is similar to its expression pattern in corresponding rat tissues. To better understand the roles of Ssp411 in male reproduction in vivo, we identified and characterized an Ssp411 expression-disrupted murine strain (Ssp411PB/PB) that was generated by piggyBac (PB) transposon insertion. We studied Ssp411-interacting proteins using proteome microarray, co-IP and GST pull-down assay.Results
Both Ssp411 mRNA and protein were detected exclusively in spermatids after step 9 during spermiogenesis in testis. Phenotypic analysis suggested that only Ssp411PB/PB males are sterile. These males have smaller testes, reduced sperm counts, decreased sperm motility and deformed spermatozoa. Microscopy analysis indicated that the manchette, a structurally reshaped sperm head, is aberrant in Ssp411PB/PB spermatids. The results of proteome microarray analysis and GST pull-down assays suggested that Ssp411 participates the ubiquitin-proteasome system by interacting with PSMC3. This has been reported to be manchette-associated and important for the head shaping of spermatids.Conclusions
Our study suggested that Ssp411 is required for spermiogenesis. It seems to play a role in sperm head shaping. The lack of Ssp411 causes sperm deformation and results in male infertility.General significance
Ssp411PB/PB mouse strain is an animal model of idiopathic oligoasthenoteratozoospermia (iOAT), and the gene may represent a therapeutic target for iOAT patients. 相似文献89.
Yuan Xu Chen Shao Xinpeng Fan Alan Warren Khaled A. S. Al-Rasheid Weibo Song Norbert Wilbert 《Polar Biology》2016,39(8):1439-1453
Antarctica is a remote and isolated biotope which makes it an ideal location for studying new and endemic species. Since there is little literature available on the diversity of ciliates in this area, a taxonomic survey of ciliates from melt-water of Collins glacier, King George Island, was carried out from January to March 2006. As a result, the morphology and infraciliature of five ciliates, including one new species, are described using live observations and silver staining: Gastronauta multistriata nov. spec., Neokeronopsis asiatica Foissner et al., 2010, Paraholosticha muscicola Kahl, 1932, Oxytricha sp., and Cyclidium glaucoma Müller, 1786. Gastronauta multistriata nov. spec. is distinguished from its congeners by the following combination of characters: cell size in vivo on average 80 × 40 μm; 5–9 kineties in left ciliary field; 18–23 kineties in right ciliary field, including 10–12 postoral kineties; 7–10 preoral kineties; dorsal brush along anterior dorsal margin, consisting of 5–8 groups of basal bodies. The only minor differences between the current population of N. asiatica and a previously described Antarctic population are the numbers of caudal cirri (6–10 vs. 8–15) and dorsal kineties (11–13 vs. 12–18). Paraholosticha sterkii is synonymised with P. muscicola. The Antarctic population of C. glaucoma corresponds well with a former population from China, the only difference being the number of kinetids in SK n (11–17 vs. 9–11). This work will contribute to the understanding of ciliate diversity in this little studied area. 相似文献
90.
Construction of a high density integrated genetic map for cucumber (Cucumis sativus L.) 总被引:1,自引:0,他引:1
Zhang WW Pan JS He HL Zhang C Li Z Zhao JL Yuan XJ Zhu LH Huang SW Cai R 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2012,124(2):249-259
The high-density consensus map was constructed based on the GY14 × PI 183967 map from an inter-subspecific cross and the extended
S94 × S06 map from an intra-subspecific cross. The consensus map was composed of 1,369 loci, including 1,152 SSR loci, 192
SRAP loci, 21 SCAR loci and one STS locus as well as three gene loci of fruit external quality traits in seven chromosomes,
and spanned 700.5 cM, of which 682.7 cM (97.5%) were covered by SSR markers. The average genetic distance and physical interval
between loci were 0.51 cM and ~268 kbp, respectively. Additionally, the physical position of the sequence-associated markers
aligned along the assembled cucumber genome sequence established a relationship between genetic maps and cucumber genome sequence
and to a great extent validated the order of markers in individual maps and consensus map. This consensus map with a high
marker density and well-ordered markers is a saturated and reliable linkage map for genetic analysis of cucumber or the Cucurbitaceae
family of plants. 相似文献