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31.
Analysis of gene function in somatic mammalian cells using small interfering RNAs 总被引:175,自引:0,他引:175
RNA interference (RNAi) is a highly conserved gene silencing mechanism that uses double-stranded RNA (dsRNA) as a signal to trigger the degradation of homologous mRNA. The mediators of sequence-specific mRNA degradation are 21- to 23-nt small interfering RNAs (siRNAs) generated by ribonuclease III cleavage from longer dsRNAs. Twenty-one-nucleotide siRNA duplexes trigger specific gene silencing in mammalian somatic cells without activation of the unspecific interferon response. Here we provide a collection of protocols for siRNA-mediated knockdown of mammalian gene expression. Because of the robustness of the siRNA knockdown technology, genomewide analysis of human gene function in cultured cells has now become possible. 相似文献
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Heidi Borgeraas Jens Kristoffer Hertel Gard Frodahl Tveitev?g Svingen Eva Ringdal Pedersen Reinhard Seifert Ottar Nyg?rd J?ran Hjelmes?th 《PloS one》2016,11(3)
Background
Asymmetric dimethylarginine (ADMA) is associated with increased risk of atherosclerotic cardiovascular disease and mortality through inhibition of nitrogen oxide (NO) synthesis. As positive correlations between serum concentrations of NO and body mass index (BMI) have been observed, we aimed to explore whether the potential associations between plasma ADMA levels and the risk of acute myocardial infarction (AMI) and mortality were modified by BMI.Methods
Multivariable Cox proportional hazard models were used to estimate the hazard ratios (HR) for AMI, cardiovascular death and all-cause mortality according to baseline plasma ADMA levels in 4122 patients with suspected stable angina pectoris. Analyses were subsequently repeated in patients with BMI below (low BMI) or above (high BMI) median.Results
A total of 2982 patients (72%) were men. Median (range) age, plasma ADMA level and BMI were 62 (21–88) years, 0.54 (0.10–1.25) μmol/L and 26.3 (18.5–54.3) kg/m2, respectively. During a mean (standard deviation) follow-up time of 4.7 (1.4) years, 337 (8%) patients suffered from an AMI, 300 (7%) died, whereof 165 (55%) due to cardiovascular disease. Each 0.1 μmol/L increment in plasma ADMA level was associated with an increased risk of AMI (HR (95% CI) 1.21 (1.08, 1.35) and cardiovascular death 1.30 (1.13, 1.49) in participants with low BMI only. Interactions were significant for AMI (p = 0.04) and CV death (p = 0.03). BMI did not modify the association between plasma ADMA levels and all-cause mortality.Conclusion
Plasma ADMA levels were associated with risk of AMI and cardiovascular death among patients with low BMI only. 相似文献33.
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Kraiczy P Hellwage J Skerka C Becker H Kirschfink M Simon MM Brade V Zipfel PF Wallich R 《The Journal of biological chemistry》2004,279(4):2421-2429
The etiologic agent of Lyme disease, Borrelia burgdorferi, is capable of circumventing the immune defense of a variety of potential vertebrate hosts. Previous work has shown that interaction of host-derived complement regulators, factor H and factor H-like protein 1 (FHL-1), with up to five complement regulator-acquiring surface proteins (CRASPs) expressed by resistant B. burgdorferi sensu lato isolates conferred complement resistance. In addition expression of CRASP-1 is directly correlated with complement resistance of Borrelia species. This work describes the functional characterization of BbCRASP-1 as the dominant factor H and FHL-1-binding protein of B. burgdorferi. The corresponding gene, zs7.a68, is located on the linear plasmid lp54 and is different from factor H-binding Erp proteins that are encoded by genes localized on circular plasmids (cp32). Deletion mutants of BbCRASP-1 were generated, and a high affinity binding site for factor H and FHL-1 was mapped to the C terminus of BbCRASP-1. Similarly, the predominant binding site of factor H and FHL-1 was localized to the short consensus repeat 7. Factor H and FHL-1 maintain their cofactor activity for factor I-mediated C3b inactivation when bound to BbCRASP-1, and factor H is up to 6-fold more efficient in mediating C3b conversion than FHL-1. In conclusion, BbCRASP-1 (i). binds the host complement regulators factor H and FHL-1 with high affinity, (ii). is the key molecule of the complement resistance of spirochetes, and (iii). is distinct from the Erp protein family. Thus, BbCRASP-1 most likely contributes to persistence of B. burgdorferi and to pathogenesis of Lyme disease. 相似文献
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Krause JC Tsibane T Tumpey TM Huffman CJ Basler CF Crowe JE 《Journal of virology》2011,85(20):10905-10908
The conserved influenza virus hemagglutinin (HA) stem domain elicits cross-reactive antibodies, but epitopes in the globular head typically elicit strain-specific responses because of the hypervariability of this region. We isolated human monoclonal antibody 5J8, which neutralized a broad spectrum of 20th century H1N1 viruses and the 2009 pandemic H1N1 virus. Fine mapping of the interaction unexpectedly revealed a novel epitope between the receptor-binding pocket and the Ca2 antigenic site on HA. This antibody exposes a new mechanism underlying broad immunity against H1N1 influenza viruses and identifies a conserved epitope that might be incorporated into engineered H1 virus vaccines. 相似文献
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Distribution and diversity of palms in a tropical biodiversity hotspot (Thailand) assessed by species distribution modeling 下载免费PDF全文
Jantrararuk Tovaranonte Anne Blach‐Overgaard Ratchada Pongsattayapipat Jens‐Christian Svenning Anders S. Barfod 《Nordic Journal of Botany》2015,33(2):214-224
Species distribution modeling has been widely used to address questions related to ecology, biogeography and species conservation on global and regional scales. Here, we study palms (Arecaceae) in a tropical biodiversity hotspot (Thailand) using species distribution modeling to assess range‐limiting factors and estimate distribution and diversity patterns based on a comprehensive compilation of occurrence records. We focused on palms as a model group due to their key‐stone importance for ecosystem functioning and socio‐economics. Different combinations of climatic, non‐climatic environmental and spatial predictors were used. The most accurate models as indicated by the ‘area under the receiver operating characteristic curve’ (AUC) statistic were those that combined all predictors. The four strongest single predictors of palm species distributions were, in decreasing order of importance, 1) latitude, 2) precipitation of driest quarter, 3) annual precipitation, and 4) minimum temperature of the coldest month, suggesting rainfall patterns and latitudinal spatial constraints as the main range determinants. Overlaying the predicted distributions revealed that potential palm hotspots are situated in the provinces of Satun and Yala in southern Thailand where vast areas remain relatively open to the discovery of new palm records and perhaps even new species. 相似文献
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