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排序方式: 共有446条查询结果,搜索用时 15 毫秒
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酪氨酸羟化酶基因载体-pCMVTH 质粒的构建及在大鼠骨骼肌内的表达 总被引:4,自引:0,他引:4
为用转基因方法治疗巴金森氏病大鼠模型,本研究采用分子克隆技术,将合成多巴胺的关键酶-酪氨酸羟化酶(TH)的基因,克隆进入以巨细胞病毒CMV为启动子的载体质粒内,经限制性内切酶定位分析证实该重组的DNA质粒的可靠性。携带TH基因的PCMVTH质粒以LIPO-FECTIN介导,在培养的原代骨骼肌细胞中高效表达。本研究为进一步用转基因的细胞植入脑内以治疗巴金森氏病打下一定基础。 相似文献
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乙肝前S2(HBVPreS2)肽段由55个氨基酸组成,其N端肽段含Th和B细胞抗原决定簇。我们将化学合成的PreS2epitope(120-145)基因与HBcAg基因不同位点进行融合,融合基因在大肠杆菌中获得表达,并对融合蛋白进行了纯化。经ELISA和Western-blot实验表明,融合蛋白具有PreS2和HBcAg两者的抗原性。此外,研究还表明,强启动子能使表达水平有一定提高。 相似文献
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Tuo Li Annika J. E. Borg Leo Krammer Rolf Breinbauer Bernd Nidetzky 《Biotechnology and bioengineering》2023,120(6):1506-1520
Polyphenolic aglycones featuring two sugars individually attached via C-glycosidic linkage (di-C-glycosides) represent a rare class of plant natural products with unique physicochemical properties and biological activities. Natural scarcity of such di-C-glycosides limits their use-inspired exploration as pharmaceutical ingredients. Here, we show a biocatalytic process technology for reaction-intensified production of the di-C-β-glucosides of two representative phenol substrates, phloretin (a natural flavonoid) and phenyl-trihydroxyacetophenone (a phenolic synthon for synthesis), from sucrose. The synthesis proceeds via an iterative two-fold C-glycosylation of the respective aglycone, supplied as inclusion complex with 2-hydroxypropyl β-cyclodextrin for enhanced water solubility of up to 50 mmol/L, catalyzed by a kumquat di-C-glycosyltransferase (di-CGT), and it uses UDP-Glc provided in situ from sucrose by a soybean sucrose synthase, with catalytic amounts (≤3 mol%) of UDP added. Time course analysis reveals the second C-glycosylation as rate-limiting (0.4–0.5 mmol/L/min) for the di-C-glucoside production. With internal supply from sucrose keeping the UDP-Glc at a constant steady-state concentration (≥50% of the UDP added) during the reaction, the di-C-glycosylation is driven to completion (≥95% yield). Contrary to the mono-C-glucoside intermediate which is stable, the di-C-glucoside requires the addition of reducing agent (10 mmol/L 2-mercaptoethanol) to prevent its decomposition during the synthesis. Both di-C-glucosides are isolated from the reaction mixtures in excellent purity (≥95%), and their expected structures are confirmed by NMR. Collectively, this study demonstrates efficient glycosyltransferase cascade reaction for flexible use in natural product di-C-β-glucoside synthesis from expedient substrates. 相似文献
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Munoz Frances M. Patel Priya A. Gao Xinghua Mei Yixiao Xia Jingsheng Gilels Sofia Hu Huijuan 《Purinergic signalling》2020,16(1):97-107
Purinergic Signalling - Astrocytes mediate a remarkable variety of cellular functions, including gliotransmitter release. Under pathological conditions, high concentrations of the purinergic... 相似文献
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肾综合征出血热(hemorrhagic fever with renal syndrome, HFRS)是一种啮齿动物传播的自然疫源性疾病, 危害严重, 已成为全球重要的公共卫生问题。本研究采用数理统计模型及小波分析方法, 对陕西省西安市鄠邑区1984-2016年HFRS的发生与鼠类、气候和经济因素的关系进行分析, 探讨气候和经济因素对HFRS发生的影响。小波分析结果表明, 该地区的HFRS暴发史可能分为两个时期, 推测每个时期具有不同的主要宿主, 在2002年褐家鼠(Rattus norvegicus)可能取代黑线姬鼠(Apodemus agrarius)成为HFRS疫源地的主要宿主。广义可加模型模拟结果表明, HFRS的发生与1984-2001年黑线姬鼠密度间存在极显著非线性效应(F2.06,9.02 = 102.415, P < 0.01), 两者间显现为正相关; 与2002-2016年的褐家鼠密度间呈正相关(F1.67,9.02 = 73.929, P < 0.01); HFRS主要宿主的这种变化可能与当地气候变化和经济发展有关: HFRS的发生与年平均温度存在极显著的非线性效应(F2.93,9.02 = 12.164, P < 0.01), 两者间呈负相关; 同样, HFRS的发生与上一年的国内生产总值(GDP)也存在显著非线性效应(F1.70,9.02 = 2.917, P < 0.05), 两者间也呈负相关。结构方程模型通过直接和间接的影响途径证明了这种转移机制, 发现温度对HFRS发生有显著的直接负向影响以及通过褐家鼠的间接正向影响; GDP对HFRS发生有直接的负向影响。本研究表明HFRS的发生与气候变化和经济发展相关, 两者均能影响HFRS的暴发, 该结论有助于今后更好地对HFRS疾病进行预防和控制。 相似文献
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Shanchao Zhang Lei Liu Ruijin Wang Houzhen Tuo Yanjun Guo Li Yi Dexin Wang Jiawei Wang 《PloS one》2013,8(12)
Human cytomegalovirus(HCMV) infection has been shown to contribute to vascular disease through the induction of angiogenesis. However, the role of microRNA in angiogenesis induced by HCMV infection remains unclear. The present study was thus designed to explore the potential effect of miR-1217 on angiogenesis and to disclose the underlying mechanism in endothelial cells. We found that HCMV infection of endothelial cells(ECs) enhanced expression of miR-217 and reduced SIRT1 and FOXO3A protein level in 24 hours post infection(hpi). Transfection of miR-217 inhibitor not only depressed cellular migration and tube formation induced by HCMV infection, but also enhanced SIRT1 and FOXO3A protein expression. Additionally, luciferase assay confirmed that miR-217 directly targeted FOXO3A mRNA 3`UTR. Furthermore, pretreatment with resveratrol depressed motility and tube formation of HCMV-infected ECs, which could be reversed by SIRT1 siRNA. Similarly, delivery of FOXO3A overexpression lentivirus suppressed proliferative rate, migration and tube formation of HCMV-infected ECs, which reversed by transfection of FOXO3A siRNA. In summary, HCMV infection of endothelial cells induces angiogenesis by both of miR-217/SIRT1 and miR-217/FOXO3A axis. 相似文献