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81.
82.
为了解锦鲤疱疹病毒中国吉林株(KHV-CJ)ORF27基因编码蛋白的结构特征和进化关系,采用镜鲤尾鳍原代细胞增殖KHV-CJ,提取其DNA,经PCR扩增,获得ORF27基因,将其克隆在pMD18-T载体中,构建重组质粒。应用生物信息学方法初步分析KHV-CJ ORF27基因的结构和功能,并与GenBank上公布的三株KHV构建系统进化树。结果显示:获得207bp的ORF27基因,编码69个氨基酸;预测ORF27编码蛋白的理论分子质量为7366.62Da,等电点为4.487;抗原表位预测显示抗原性良好;编码蛋白存在1个N-糖基化位点、4个O糖基化位点和5个磷酸化位点;系统进化树分析表明与锦鲤疱疹病毒美国株(KHV-U)属同一分支。 相似文献
83.
将禽白血病病毒p27基因克隆并构建重组表达质粒pET28a-p27,在大肠杆菌BL21中经IP TG诱导后产生可溶性表达蛋白。表达的蛋白用Western blot进行活性检测,其能与辣根过氧化物酶标记的兔抗p27抗体发生特异性反应;采用金属螯和层析对表达蛋白进行纯化,其纯度约为95%;用纯化的表达蛋白p27免疫家兔制备抗血清,ELISA抗体效价可达1∶25600。研究结果表明,大肠杆菌表达的p27蛋白具有良好的抗原性,可以替代纯化病毒蛋白用于禽白血病病毒检测。 相似文献
84.
采用细胞培养法增殖禽白血病/肉瘤病毒(ALV/RSV)A亚型,然后对其进行超速离心浓缩及蔗糖密度梯度离心纯化病毒,再采用电泳分离与电洗脱收集相结合的方法纯化p27蛋白。经检测,提取的p27蛋白具有良好的抗原性与免疫原性。用该蛋白免疫家兔制备的兔抗p27抗体,经ELISA方法检测其效价可达1:6400以上。 相似文献
85.
Catherine Albertini Pierre Leroux 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(7):723-733
Botrytis cinerea (anamorph of Botryotinia fuckeliana) is a filamentous ascomycete that causes grey mould on grapevine. We had previously described two distinct populations, named HydR1 and non-HydR1, that comprise two distinct genetic entities based on genetic polymorphism, natural resistance towards the fungicide hydroxyanilide fenhexamid, and vegetative incompatibility between them. Here, we used PCR to isolate the 3-keto reductase gene ERG27 by virtue of sequence homology with Saccharomyces cerevisiae ERG27. The gene product was longer than the yeast's enzyme but possessed the main characteristic features of reductases. It displayed striking homology with mammalian 17-HSD7, therefore confirming the hypothesis of a common function between Erg27p like protein and 17-HSD7 in sterol biosynthesis (i.e. cholesterol, ergosterol). On the other hand, we analysed the polymorphism of the B. cinerea gene product and found a dozen of amino-acid differences between strains of HydR1 and non-HydR1 types that could underlie HydR1 natural resistance to fenhexamid. First, this polymorphism analysis showed that HydR1 strains form a homogeneous group distinct from the non-HydR1 group of strains. These results support our hypothesis that HydR1 and non-HydR1 strains constitute two different species. Second, Erg27p like protein sequence analysis showed that a high resistant phenotype to fenhexamid, HydR3, found in treated populations of non-HydR1 strains, had two mutations (usually found in mammalian 17-HSD7) that could be useful as population markers. 相似文献
86.
J. J. Geng Y. H. Shen F. Y. Yang K. Li 《The Journal of Horticultural Science and Biotechnology》2016,91(2):203-209
Papaya (Carica papaya L.) fruit have a short shelf-life due to rapid ripening induced by ethylene, which usually results in a high percentage of product loss. However, little is known about the genetic mechanism of ripening and the attributes of fruit quality. Ubiquitin (UBQ) proteins have received increasing attention because they play important roles in response to ripening and in regulating certain developmental processes in plants. In the present study, three genes encoding UBQ proteins, CpUBI1, CpUBI2, and CpUBI3, were isolated from papaya fruit. The lengths of the cDNAs of CpUBI1, CpUBI2, and CpUBI3 were 1,485 bp, 1,642 bp, and 529 bp, encoding 306, 308, and 156 predicted amino acids, respectively. Sequence analysis demonstrated that the CpUBI1 and CpUBI2 proteins contained four consecutive structural domains of the UBQ superfamily, while CpUBI3 contained a ribosomal domain structure of the S27a superfamily. RT-qPCR analysis demonstrated that ethephon treatment increased CpUBI gene expression significantly, compared to 1-methylcyclopropene (1-MCP) treatment and the untreated controls. Levels of CpUBI1 and CpUBI2 gene expression were significantly higher than CpUBI3. These results suggested that CpUBI1, CpUBI2, and CpUBI3 might have different roles during papaya fruit ripening and softening. 相似文献
87.
热休克蛋白27(Hsp27)是一种结构上高度保守的小热休克应激蛋白,具有多重生物学功能,在调节蛋白质稳态以维持细胞骨架有至关重要的作用。近年来,热休克蛋白(HSPs)在疾病预防和药物开发中取得较大突破,有研究发现在许多病毒感染宿主细胞后,细胞中Hsp27的表达会发生显著变化,表明Hsp27与病毒感染之间有密切关联,但Hsp27在病毒感染中的应用尚不成熟。论文主要介绍病毒感染宿主细胞后,Hsp27通过抑制细胞的自噬、凋亡、干扰素形成及维持病毒复制等发挥其在病毒感染中作用,此方面研究为以HSPs为靶点研发抗病毒药物提供了新思路。 相似文献
88.
GUO Gui-xian CHENG Chuan-fang GU Jie-lei ZHOU Wen-yi TANG Xiao-yan ZHONG Yun CHEN Yan-fang LIU Shi-ming 《园艺学报》2018,34(12):2113-2119
AIM:To investigate the effect of inhibiting the release of exosomes on the biological characteristics of bone marrow mesenchymal stem cells (BM-MSCs) and the underlying mechanisms. METHODS:The exosome releasing-deficient mouse model was constructed by knockout of Rab27a using TALEN technique. The BM-MSCs were isolated and cultured. The exosomes were extracted from the culture medium using total exosome isolation kit and quantified by nanoparticle tracking analysis (NTA). The size and morphology of the exosomes were observed under transmission electron microscope. To evaluate the proliferation ability of BM-MSCs, the BM-MSCs were labeled with 5-ethynyl-2'-deoxyuridine (EdU) and the expression level of proliferating cell nuclear antigen (PCNA) was determined by Western blot. Moreover, hypoxia tolerance of BM-MSCs in vitro was evaluated via TUNEL staining and MTS assay. RESULTS:The count of exosomes released by BM-MSCs isolated from Rab27a knockout mice was significantly reduced. Inhibition of exosome release resulted in decreases in the viability of the BM-MSCs and their resistance to hypoxia. CONCLUSION:Inhibition of exosome release from the BM-MSCs results in significantly decreased proliferation ability and resistance to hypoxia. 相似文献
89.
Interactions between pulp fibers and aluminum compounds in pulp suspensions were studied using fibrous cellulose (FC) and fibrous carboxymethylcellulose (FCMC) powders as models of pulp fibers by X-ray fluorescence analysis and27Al nuclear magnetic resonance. When deionized water was used at pH 4–5, water-soluble cationic aluminum species (Al3+, aluminum oligomer, and polyaluminum species) were adsorbed on the solid FCMC, forming carboxylic acid aluminum salts by cation exchange. The formation of these nondissociated pulp-COOAl type structures in paper sheets may contribute to some decreases in hydrophilic property. On the other hand, the water-soluble cationic aluminum species had nearly no interactions with hydroxyl groups of solid cellulose in the suspensions at pH 4–5. When tap water was used at pH 5–7, some aluminum components were retained on not only FCMC but also the FC sample. Probably, water-insoluble Al(OH)3 flocs are formed in the suspensions at pH 5–7 and retained on the FC sample by simple filtration effect. Therefore, two mechanisms of the aluminum retention (i.e., electrostatic interactions and a simple filtration effect) may exist between pulp fibers and aluminum components in the practical papermaking process.This research was presented in part at the 5th annual meeting of the Japan Cellulose Society, Kyoto, July 1998 相似文献
90.
为寻求抗ALV—J感染雏鸡用药物,通过CCK-8试剂盒检测细胞活力以确定利巴韦林、病毒灵和黄芪多糖对DFl细胞的最大安全浓度,用ELISA试剂盒检测ALV—JP27抗原表达量,以确定上述抗病毒药物体外抗ALV—J活性。结果显示,利巴韦林、病毒灵和黄芪多糖对DFl细胞的最大安全浓度分别为0.0391、0.0625和0.0625mg/mL。在安全范围内,利巴韦林对P27的表达量具有明显的抑制作用,黄芪多糖在0.0039~O.0625mg/mL安全浓度范围内明显抑制P27的表达,病毒灵在安全范围内对P27的表达基本上没有抑制作用。试验表明,利巴韦林和黄芪多糖对ALV-J的增殖有明显的抑制作用,有望作为该病净化的辅助性防制药物。 相似文献