全文获取类型
收费全文 | 67篇 |
免费 | 10篇 |
国内免费 | 2篇 |
出版年
2023年 | 1篇 |
2022年 | 1篇 |
2020年 | 1篇 |
2019年 | 1篇 |
2014年 | 2篇 |
2011年 | 3篇 |
2010年 | 4篇 |
2009年 | 3篇 |
2008年 | 2篇 |
2007年 | 1篇 |
2005年 | 1篇 |
2004年 | 3篇 |
2003年 | 3篇 |
2002年 | 1篇 |
2001年 | 6篇 |
2000年 | 5篇 |
1999年 | 4篇 |
1998年 | 3篇 |
1996年 | 3篇 |
1995年 | 1篇 |
1992年 | 4篇 |
1991年 | 1篇 |
1990年 | 1篇 |
1988年 | 1篇 |
1987年 | 1篇 |
1985年 | 1篇 |
1983年 | 1篇 |
1979年 | 2篇 |
1978年 | 3篇 |
1977年 | 4篇 |
1976年 | 3篇 |
1975年 | 2篇 |
1973年 | 1篇 |
1967年 | 1篇 |
1966年 | 1篇 |
1965年 | 1篇 |
1964年 | 1篇 |
1960年 | 1篇 |
排序方式: 共有79条查询结果,搜索用时 15 毫秒
41.
拼接信号序列单碱基变异提高马传染性贫血病毒mRNA拼接效率 总被引:1,自引:0,他引:1
分别以马传染性贫血(马传贫)驴强毒(D—A EIAV)RNA和马传贫驴白细胞弱毒疫苗(DLA EIAV)RNA为模板,利用RT—PCR的方法,克隆到马传贫强、弱毒株基因组外显子2及其下游的核苷酸序列。然后将报告基因CAT插入到EIAV内含子2env阅读框架中,构成CAT拼接报告系统。同时在强毒株重组表达质粒的基础上,将其外显子-3上游拼接受体位点的核苷酸序列CAG突变为弱毒株相应位置的核苷酸序列TAG,得到强毒单核苷酸突变株重组表达质粒。用构建的3个重组表达质粒DNA转染驴血白细胞,ELISA检测转染细胞CAT浓度。结果表明:EIAV强毒株重组表达质粒中CAT蛋白表达量最高,EIAV强毒株重组表达质粒次之,EIAV强毒突变株重组表达质粒最低。由于CAT基因被插入于各重组质粒中的EIAV内含子-2里,EIAV外显子-2、3之间的拼接可导致该基因的删除,因而其拼接效率低于EIAVmRNA外显子-2、3之间的拼接效率。实验数据表明,EIAV SA2拼接信号序列单碱基变异提高了SD2-SA2拼接效率;D—AEIAV SA2-SD2拼接效率比DLA EIAV相应位点拼接效率高。 相似文献
42.
结核分枝杆菌可视化抗体检测蛋白芯片的制备 总被引:1,自引:0,他引:1
目的:利用克隆表达的7种结核分枝杆菌优势表位抗原,建立可视化抗体检测蛋白芯片,用于结核病辅助诊断。方法:将7种结核分枝杆菌优势表位抗原,即38kD、ESAT-6、CFP10、MPT64、Mtb8、Mtb8.4和Mtb16.3点于修饰的基片上,制备可检测7种结核抗体的多靶点蛋白微阵列,建立免疫金银染色检测系统;使用该芯片对48例临床结核病患者血液样品进行检测,并与“金标准”痰涂片(48例)和痰培养(其中的29例)检测结果进行比较,分析其敏感性;对30名献血员血液样品进行检测,分析其特异性。结果:可视化抗体检测蛋白芯片的敏感性分别为98.5%和96.6%,而痰涂片和痰培养检测方法的敏感性分别为35.4%和48.3%;可视化抗体检测蛋白芯片的特异性为93.3%。结论:建立的结核分枝杆菌可视化抗体检测蛋白芯片检测敏感性显著高于痰涂片和痰培养方法,可用于结核病的临床辅助诊断,提高痰涂片和痰培养假阴性的检出率。 相似文献
43.
Background
Species are fundamental units in biology, yet much debate exists surrounding how we should delineate species in nature. Species discovery now requires the use of separate, corroborating datasets to quantify independently evolving lineages and test species criteria. However, the complexity of the speciation process has ushered in a need to infuse studies with new tools capable of aiding in species delineation. We suggest that model-based assignment tests are one such tool. This method circumvents constraints with traditional population genetic analyses and provides a novel means of describing cryptic and complex diversity in natural systems. Using toad-headed agamas of the Phrynocephalus vlangalii complex as a case study, we apply model-based assignment tests to microsatellite DNA data to test whether P. putjatia, a controversial species that closely resembles P. vlangalii morphologically, represents a valid species. Mitochondrial DNA and geographic data are also included to corroborate the assignment test results. 相似文献44.
Pletnev V. Z. Goryacheva E. A. Tsygannik I. N. Nesmeyanov V. A. Pletnev S. V. Pangborn W. Daux W. 《Russian Journal of Bioorganic Chemistry》2004,30(5):417-420
The three-dimensional structure of the antigen-binding fragment of a monoclonal antibody to human interleukin-2 in a new crystal form (space group P212121; unit cell parameters: a = 42.82 Å, b = 90.68 Å, and c = 139.82 Å) was determined by the X-ray molecular replacement method at the resolution of 2.7 Å. The protein folding and the stereochemistry of its antigen-binding site were comparatively analyzed. 相似文献
45.
46.
Crystal structure of bovine duodenase, a serine protease, with dual trypsin and chymotrypsin-like specificities 总被引:2,自引:0,他引:2
The three-dimensional structure of duodenase, a serine protease from bovine duodenum mucosa, has been determined at 2.4A resolution. The enzyme, which has both trypsin-like and chymotrypsin-like activities, most closely resembles human cathepsin G with which it shares 57% sequence identity and similar specificity. The catalytic Ser195 in duodenase adopts the energetically favored conformation typical of serine proteinases and unlike the strained state typical of lipase/esterases. Of several waters in the active site of duodenase, the one associated with Ser214 is found in all serine proteinases and most lipase/esterases. The conservation of the Ser214 residue in serine proteinase, its presence in the active site, and participation in a hydrogen water network involving the catalytic triad (His57, Asp107, and Ser195) argues for its having an important role in the mechanism of action. It may be referred to as a fourth member of the catalytic triad. Duodenase is one of a growing family of enzymes that possesses trypsin-like and chymotrypsin-like activity. Not long ago, these activities were considered to be mutually exclusive. Computer modeling reveals that the S1 subsite of duodenase has structural features compatible with effective accommodation of P1 residues typical of trypsin (Arg/Lys) and chymotrypsin (Tyr/Phe) substrates. The determination of structural features associated with functional variation in the enzyme family may permit design of enzymes with a specific ratio of trypsin and chymotrypsin activities. 相似文献
47.
The effective population size of Anopheles gambiae in Kenya: implications for population structure 总被引:4,自引:0,他引:4
We estimated current and long-term effective population size (Ne) of two
Anopheles gambiae (savanna cytotype) populations in Kenya. Temporal
variation at nine microsatellite loci in each population sampled 7 and 9
years apart and genetic diversity in each sample were analyzed to answer
the following questions. (1) Do bottlenecks occur in Kenyan populations of
A. gambiae? (2) How variable are different populations with respect to
their current and long-term Ne values? (3) What are the implications of
these results on population structure and history? The estimates of Ne of
Asembo and Jego were 6,359 and 4,258, respectively, and the lower 95%
limits were 2,455 and 1,669, respectively. Thus, despite the typical
observation of low density at the village level during the dry season,
large populations are maintained annually. Large current Ne is consistent
with previous studies showing low differentiation across the continent,
especially under Wright's isolation-by-distance model. Current Ne in Asembo
was 1.5-fold higher than in Jego, but this difference was not significant.
Long-term Ne in Asembo (22,667) was 2.9-fold higher than that in Jego
(7,855) based on the stepwise mutation model. The difference between
populations was significant at both time points regardless of whether
long-term Ne values were calculated based on the stepwise mutation model or
the infinite-alleles model. Heterozygosity in Jego declined significantly
between 1987 (59%) and 1996 (54%), whereas heterozygosity in Asembo was
stable (66%-65%). Despite the relatively high and significant
differentiation between Asembo and Jego (FST = 0.072-0.10, RST = 0.037-
0.038), all alleles in Jego were found in Asembo but not vice versa. All of
these findings suggest that lower Ne in Jego magnifies differentiation
between the two populations. The long-term Ne was biased downward, because
its calculation was based on an upper bound estimate of microsatellite
mutation rate. Ne values based on mtDNA and allozymes were an order of
magnitude higher. Long-term Ne therefore, is probably measured in hundreds
of thousands and hence does not support a recent expansion of this species
from a small population.
相似文献
48.
The linear pentadecapeptide antibiotic gramicidin D is a heterogeneous mixture of six components. Precise refinements of three-dimensional structures of naturally occurring gramicidin D in crystals obtained from methanol, ethanol, and n-propanol demonstrate the unexpected presence of stable left-handed antiparallel double-helical heterodimers that vary with the crystallization solvent. The side chains of Trp residues in the three structures exhibit sequence-specific patterns of conformational preference. Tyr substitution for Trp at position 11 appears to favor beta ribbon formation and stabilization of the antiparallel double helix that acts as a template for gramicidin folding and nucleation of different crystal forms. The fact that a minor component in a heterogeneous mixture influences aggregation and crystal nucleation has potential applications to other systems in which anomalous behavior is exhibited by aggregation of apparently homogeneous materials, such as the enigmatic behavior of prion proteins. 相似文献
49.
Folylpolyglutamate synthetase (FPGS) from Lactobacillus casei has been crystallized with polyethylene glycol and acetate buffer at pH 5.0. The enzyme was obtained from Escherichia coli strain SF4 harboring the L. casei FPGS chromosomal gene on a pEMBL vector (pGT3-8.1). Crystals of the enzyme were obtained which diffract to 2.6 A resolution. The crystals are monoclinic, space group P2(1), with unit cell dimensions of a = 54.07 A, b = 45.83 A, c = 84.37 A and beta = 107.92 degrees. A unit cell contains one molecule of the 43,000 Da enzyme per asymmetric unit. A complete X-ray data set on the native crystals has been collected. 相似文献
50.
The plasmalemma and hyaline ectoplasm together constitute the sensory and motor organ of macrophages. The purpose of this study was to isolate this cell fraction in order to analyze it biochemically and functionally. Brief sonification of warmed rabbit lung macrophages caused release of heterodisperse hyaline blebs and filopodia, which were easily collected by differential centrifugation. Viewed in the electron microscope, these structures consisted of membrane-bounded sacs principally containing actin filaments. Some contained secondary lysosomes. They were enriched threefold over whole cell homogenates in specific adenylate cyclase activity and in trichloroacetic-acid-precipitable (125)I when derived from cells labeled with 125(I) by means of a lactoperoxidase-catalyzed reaction. These markers were found to have identical isopycnic densitites when macrophage homogenates were subjected to sedimentation in a focusing sucrose density gradient system, and these markers had densities distinct from those of other cytoplasmic organelles. These markers were therefore assumed to be associated with macrophage plasma membranes. The specific β- glucuronidase activity of the bleb fraction was similar to that of homogenates, but the blebs had considerably lower specific succinic dehydrogenase activity and RNA content, and DNA was undetectable. Electrophoresis of blebs solubilized in sodium dodecyl sulfate on polyacrylamide gels revealed polypeptides co-migrating with macrophage actin-binding protein, myosin, and actin; blebs also had EDTA-activated adenosine triphosphatase activity characteristic of myosin. The concentrations of actin-binding protein and myosin were higher in blebs than in cells or cytoplasmic extracts, whereas actin concentrations were similar (relative to extracts) or only slightly greater (than in cells). Blebs and intact cells had high lactate dehydrogenase activities in the presence but not the absence of Triton X-100. Blebs and cells oxidased 1-[(14)C]glucose, and the rate of glucose oxidation was increased substantially in the presence of latex beads. We conclude that intact sacs of plasmalemma encasing contractile proteins and cytoplasmic enzymes can be isolated from macrophages. They are enriched in myosin and actin-binding protein, indicating that the contractile apparatus is regulated in the cell periphery. These structures have the capacity to respond to environmental signals. We suggest the name "podosomes" for them because of their resemblance to macrophage pseudopodia. We propose that podosome formation results from rapid dissolution of the cortical gel when the membrane is in an actively extended configuration. 相似文献