全文获取类型
收费全文 | 3669篇 |
免费 | 307篇 |
国内免费 | 69篇 |
出版年
2023年 | 12篇 |
2022年 | 39篇 |
2021年 | 79篇 |
2020年 | 38篇 |
2019年 | 52篇 |
2018年 | 60篇 |
2017年 | 66篇 |
2016年 | 96篇 |
2015年 | 121篇 |
2014年 | 157篇 |
2013年 | 188篇 |
2012年 | 243篇 |
2011年 | 232篇 |
2010年 | 162篇 |
2009年 | 144篇 |
2008年 | 188篇 |
2007年 | 220篇 |
2006年 | 204篇 |
2005年 | 177篇 |
2004年 | 161篇 |
2003年 | 164篇 |
2002年 | 156篇 |
2001年 | 70篇 |
2000年 | 65篇 |
1999年 | 83篇 |
1998年 | 35篇 |
1997年 | 47篇 |
1996年 | 44篇 |
1995年 | 33篇 |
1994年 | 35篇 |
1993年 | 27篇 |
1992年 | 60篇 |
1991年 | 53篇 |
1990年 | 61篇 |
1989年 | 57篇 |
1988年 | 56篇 |
1987年 | 41篇 |
1986年 | 44篇 |
1985年 | 34篇 |
1984年 | 33篇 |
1983年 | 26篇 |
1982年 | 32篇 |
1981年 | 21篇 |
1980年 | 18篇 |
1979年 | 22篇 |
1978年 | 13篇 |
1977年 | 12篇 |
1976年 | 11篇 |
1975年 | 9篇 |
1973年 | 11篇 |
排序方式: 共有4045条查询结果,搜索用时 15 毫秒
81.
Kan Wang Paul Drayton Bronwyn Frame Jim Dunwell John Thompson 《In vitro cellular & developmental biology. Plant》1995,31(2):101-104
A number of different methods, involving direct DNA delivery are now available for plant transformation. Here we review the most recently developed technique which involves the mixing of silicon carbide whiskers with plant cells and plasmid DNA. Fertile transgenic plants have now been produced using whisker-mediated transformation, and this method can now be considered as a simple, inexpensive alternative for plant transformation. A brief review on transformation of animal cells andChlamydomonas using whiskers technology is also included. 相似文献
82.
Jeffrey S. Rubin Donald P. Bottaro Marcio Chedid Toru Miki Dina Ron Hyae-Gyeong Cheon William G. Taylor Emma Fortney Hiromi Sakata Paul W. Finch William J. LaRochelle 《Cell biology international》1995,19(5):399-411
Keratinocyte growth factor (KGF) is a member of the heparin-binding fibroblast growth factor family (FGF-7) with a distinctive pattern of target-cell specificity. Studies performed in cell culture suggested that KGF was mitogenically active only on epithelial cells, albeit from a variety of tissues. In contrast, KGF was produced solely by cells of mesenchymal origin, leading to the hypothesis that it might function as a paracrine mediator of mesenchymal-epithelial communication. Biochemical analysis and molecular cloning established that the KGF receptor (KGFR) was a tyrosine kinase isoform encoded by the fgfr-2 gene. Many detailed investigations of KGF and KGFR expression in whole tissue and cell lines largely substantiated the pattern initially perceived in vitro of mesenchymal and epithelial distribution, respectively. Moreover, functional assays in organ culture and in vivo and studies of KGF regulation by sex sterorid hormones reinforced the idea that KGF acts predominantly on epithelial cells to elicit a variety of responses including proliferation, migration and morphogenesis. 相似文献
83.
A novel oncogene, ost, encodes a guanine nucleotide exchange factor that potentially links Rho and Rac signaling pathways. 总被引:19,自引:5,他引:14
下载免费PDF全文
![点击此处可从《The EMBO journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Transfection of NIH3T3 cells with an osteosarcoma expression cDNA library led to the appearance of foci of morphologically transformed cells which were found to harbor a novel oncogene, ost. The ost product was activated by truncation of the N-terminal domain of the ost proto-oncogene and was highly tumorigenic in nude mouse assays. The proto-ost cDNA, isolated subsequently, encodes a predicted protein of 100 kDa containing DH (Db1 homology) and PH (pleckstrin homology) domains. Ost is mainly phosphorylated on serine and localized in the cytoplasm. Purified Ost protein catalyzed guanine nucleotide exchange on RhoA and Cdc42 among the Rho and Ras family members tested, indicating that Ost can activate these small GTP-binding proteins. Ost did not detectably associate with RhoA or Cdc42, but interacted specifically with the GTP-bound form of Rac1, suggesting that Ost can function as an effector of Rac1. These results suggest that Ost is a critical regulatory component which links pathways that signal through Rac1, RhoA and Cdc42. Of the tissues examined, expression of ost was the highest in brain and could be localized to neurons and alpha-tanycytes, suggesting that Ost may participate in axonal transport in these specialized cells. 相似文献
84.
Yasuhiro Iwao Akiko Miki Michiko Kobayashi Kazuo Onitake 《Development, growth & differentiation》1994,36(5):469-479
An extract obtained from Cynops sperm induced the activation of both Cynops and Xenopus eggs with accompanying changes in the potential of the egg membrane that were quite similar to those caused by the Cynops sperm. The activation-inducing properties of the extract were abolished by treatment with proteinase K or by heating (60°C, 15 min) and were associated with a protease activity against peptidyl Arg-MCA substrates. The activation of Xenopus eggs by the extract was inhibited by those substrates, or by protease inhibitors, aprotinin or leupeptin. The protease activity was localized in the acrosomal region of Cynops sperm. The activation of Xenopus eggs by the extract was prevented when the exterior concentration of Ca2+ ions, [Ca2+ ]0 , was reduced to 1.5 μM, but it was enhanced when [Ca2+ ]0 was increased to 340 μM. The activation of Xenopus eggs by the extract was not affected by positive clamping when [Ca2+ ]0 was 340 μM. These results suggest that the sperm extract contains a protease that causes an increase in the influx of Ca2+ ions that results in voltage-insensitive activation of the egg. 相似文献
85.
Expression of the BnmNAP subfamily of napin genes coincides with the induction of Brassica microspore embryogenesis 总被引:3,自引:0,他引:3
Kim A. Boutilier María-Jesús Ginés Janice M. DeMoor Bin Huang Chris L. Baszczynski V. N. Iyer Brian L. Miki 《Plant molecular biology》1994,26(6):1711-1723
Brassica napus cv. Topas microspores can be diverted from pollen development toward haploid embryo formation in culture by subjecting them to a heat stress treatment. We show that this switch in developmental pathways is accompanied by the induction of high levels of napin seed storage protein gene expression. Changes in the plant growth or microspore culture conditions were not by themselves sufficient to induce napin gene expression. Specific members of the napin multigene family were cloned from a cDNA library prepared from microspores that had been induced to undergo embryogenesis. The majority of napin clones represented three members (BnmNAP2, BnmNAP3 and BnmNAP4) that, along with a previously isolated napin genomic clone (BngNAP1), constitute the highly conserved BnmNAP subfamily of napin genes. Both RNA gel blot analysis, using a subfamily-specific probe, and histochemical analysis of transgenic plants expressing a BngNAP1 promoter--glucuronidase gene fusion demonstrated that the BnmNAP subfamily is expressed in embryogenic microspores as well as during subsequent stages of microsporic embryo development. 相似文献
86.
T Suzuki M Murakami N Onai E Fukuda Y Hashimoto M H Sonobe T Kameda M Ichinose K Miki H Iba 《Journal of virology》1994,68(6):3527-3535
87.
Kan Shida Kotaro Takamizawa Masato Nagaoka Takao Tsuji Toshiaki Osawa 《Microbiology and immunology》1994,38(4):273-279
The binding of Escherichia coli heat-labile enterotoxin (LT) type I to glycosylated proteins with lactose (Galβ1-4Glc) by amino carbonyl reaction was studied by the Western blot assay and by the microtiter well binding assay. LT bound to a lactose-α-lactalbumin amino carbonyl product (Lac-LA), whereas cholera toxin did not. The binding ability of Lac-LA was abolished by β-galactosidase treatment, indicating that the terminal galactose is essential for the binding of LT. The binding of LT to Lac-LA was inhibited by galactose and lactose, and most effectively inhibited by lactulose (Galβ1-4Fru), which is a structural analog of the Amadori rearrangement product of the amino carbonyl reaction between lactose and an ε-amino group of a lysine residue (lactuloselysine). The results suggest that LT recognizes the portion of lactuloselysine in Lac-LA. LT also bound to a melibiose (Galα1-6Glc)-α-lactalbumin amino carbonyl product (Mel-LA), but the binding ability of Mel-LA was weaker than that of Lac-LA, suggesting that the β1-4 linked terminal galactose is dispensable but preferable for the binding. Furthermore, LT bound to the amino carbonyl products of lactose with β-lactoglobulin, caseins, bovine serum albumin, and ovalbumin. These results indicate that LT binds to the amino carbonyl products between proteins and sugars containing the terminal galactose, such as lactose. 相似文献
88.
Hideyoshi Yoshida Miki Shirakihara Akira Takemura Kunio Shirakihara 《Marine Mammal Science》1994,10(3):266-282
Development, sexual dimorphism, and individual variation were examined in the skeleton of the finless porpoise in the coastal waters of western Kyushu, Japan. Skulls ceased growing by 4 yr. Postcranial skeletons ceased increasing in size at an age older than 11 yr. The finless porpoise was estimated to attain cranial maturity by 4 yr and physical maturity at 14–23 yr. Sexual dimorphism was not detectable in most of the cranial characters but was detected in more than half of the postcranial characters. Females tended to show larger values of postcranial characters. The shape of the pelvic bone was obviously different between males and females. Thus, a discriminant function was proposed to determine sex using measurements of this bone. Individual variation was greatest in the feeding apparatus such as length of the rostrum, and least in the braincase. 相似文献
89.
Nakazawa Miki; Hayashi Hidenori; Yoshida Yuhji; Manabe Katsushi 《Plant & cell physiology》1993,34(1):83-91
Peptide fragments were obtained by limited proteolysis withtrypsin and Staphylococcus aureus V 8 protease from either thePR or the PFR form of 121-kDa phytochrome purified from etiolatedpea (Pisum sativum L.) shoots. Patterns of bands after polyacrylamidegel electrophoresis in the presence of SDS of the digests weredifferent, with some bands appearing preferentially when thedigestions were carried out with the PR or the PFR form. Amino-terminalsequences of the fragments were analyzed to determine the exactlocations of the amino-termini of the fragments within the aminoacid sequence of the apoprotein of pea phytochrome. The aminoacid compositions of some of the sequenced fragments were determinedin order to confirm the carboxy-terminal amino acids. Threecleavage regions were identified as kinetically favored sitesof cleavage of PFR (Arg-746 to Lys-752, around Glu-877 and aroundArg-1010), whereas only one was identified for PR (Glu-38 toArg-62). Regions of Glu-255, Arg-383, Arg-583 to Glu-620 andLys-1093 to Glu-1115 were also identified as potential sitesof proteolytic cleavage in both forms of the phytochrome. Othercleavage sites, the specificities of which have not yet beendetermined, are Glu-404, Glu-695 and Lys-1045. Surface-exposed parts of phytochrome in the PR and PFR formsare discussed. (Received June 13, 1992; Accepted October 27, 1992) 相似文献
90.
Ishimori H Saeki K Inai M Nagao Y Itasaka J Miki Y Seike N Kainuma H 《Theriogenology》1993,40(2):427-433
The influence of equilibration time before vitrification on the viability of vitrified morula- to blastocyst-stage bovine embryos and in vivo viability of vitrified embryos following transfer to recipients were investigated. In experiment 1, the embryos were exposed to an equilibration solution (50% VSED) containing 12.5% v/v ethylene glycol and 12.5% v/v dimethyl sulfoxide in modified Dulbecco's phosphate buffered saline with 4 mg/ml BSA (m-PBS) for 1, 2 and 5 minutes at room temperature (22 to 24 degrees C). The embryos were then placed in 15mul vitrification solution (VSED) consisting of 25% v/v ethylene glycol and 25% v/v dimethyl sulfoxide in m-PBS and were loaded into 0.25 ml plastic straws at room temperature. After 30 seconds, the straws were placed in liquid nitrogen (LN(2)) vapor for 2 minutes, plunged and stored in LN(2). To thaw, the straws were warmed in water at 20 degrees C for 15 seconds and the contents of the straws were expelled into a plastic dish. The embryos were diluted in 0.5 M sucrose + m-PBS for 5 minutes and were cultured in TCM-199 supplemented with bovine oviductal epithelial tissue. Viability of the embryos was assessed by the forming or reforming of the blastocoele after 24 hours of culture. High in vitro survival rates (73 approximately 90%) of vitrified embryos were obtained after 1 and 2 minute equilibrations, but was reduced (P<0.05) after 5 minute equilibration. In Experiment 2, morula- to blastocyst-stage embryos were vitrified after 1 minute equilibration in 50% VSED and 30 seconds of exposure to VSED. The vitrified-warmed embryos were transferred to recipient heifers at 7 days after estrus (1 embryo per recipient). Five (38%) of 13 (40%) of 10 recipients that had received blastocysts were diagnosed as pregnant using ultrasonography 60 days following transfer. 相似文献