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SARS-CoV是引起严重急性呼吸道综合症(SARS)的病原体.更多地了解SARS-CoV的基因组、蛋白结构以及它与其它冠状病毒的关系,将有助于SARS疾病的防治. 相似文献
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The ORF4a protein of human coronavirus 229E functions as a viroporin that regulates viral production
Ronghua Zhang Kai Wang Wei Lv Wenjing Yu Shiqi Xie Ke Xu Wolfgang Schwarz Sidong Xiong Bing Sun 《生物化学与生物物理学报:生物膜》2014
In addition to a set of canonical genes, coronaviruses encode additional accessory proteins. A locus located between the spike and envelope genes is conserved in all coronaviruses and contains a complete or truncated open reading frame (ORF). Previously, we demonstrated that this locus, which contains the gene for accessory protein 3a from severe acute respiratory syndrome coronavirus (SARS-CoV), encodes a protein that forms ion channels and regulates virus release. In the current study, we explored whether the ORF4a protein of HCoV-229E has similar functions. Our findings revealed that the ORF4a proteins were expressed in infected cells and localized at the endoplasmic reticulum/Golgi intermediate compartment (ERGIC). The ORF4a proteins formed homo-oligomers through disulfide bridges and possessed ion channel activity in both Xenopus oocytes and yeast. Based on the measurement of conductance to different monovalent cations, the ORF4a was suggested to form a non-selective channel for monovalent cations, although Li+ partially reduced the inward current. Furthermore, viral production decreased when the ORF4a protein expression was suppressed by siRNA in infected cells. Collectively, this evidence indicates that the HCoV-229E ORF4a protein is functionally analogous to the SARS-CoV 3a protein, which also acts as a viroporin that regulates virus production. This article is part of a Special Issue entitled: Viral Membrane Proteins — Channels for Cellular Networking. 相似文献
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Sheng-Fan Wang Sung-Pin Tseng Chia-Hung Yen Jyh-Yuan Yang Ching-Han Tsao Chun-Wei Shen Kuan-Hsuan Chen Fu-Tong Liu Wu-Tse Liu Yi-Ming Arthur Chen Jason C. Huang 《Biochemical and biophysical research communications》2014
The severe acute respiratory syndrome coronavirus (SARS-CoV) still carries the potential for reemergence, therefore efforts are being made to create a vaccine as a prophylactic strategy for control and prevention. Antibody-dependent enhancement (ADE) is a mechanism through which dengue viruses, feline coronaviruses, and HIV viruses take advantage of anti-viral humoral immune responses to infect host target cells. Here we describe our observations of SARS-CoV using ADE to enhance the infectivity of a HL-CZ human promonocyte cell line. Quantitative-PCR and immunofluorescence staining results indicate that SARS-CoV is capable of replication in HL-CZ cells, and of displaying virus-induced cytopathic effects and increased levels of TNF-α, IL-4 and IL-6 two days post-infection. According to flow cytometry data, the HL-CZ cells also expressed angiotensin converting enzyme 2 (ACE2, a SARS-CoV receptor) and higher levels of the FcγRII receptor. We found that higher concentrations of anti-sera against SARS-CoV neutralized SARS-CoV infection, while highly diluted anti-sera significantly increased SARS-CoV infection and induced higher levels of apoptosis. Results from infectivity assays indicate that SARS-CoV ADE is primarily mediated by diluted antibodies against envelope spike proteins rather than nucleocapsid proteins. We also generated monoclonal antibodies against SARS-CoV spike proteins and observed that most of them promoted SARS-CoV infection. Combined, our results suggest that antibodies against SARS-CoV spike proteins may trigger ADE effects. The data raise new questions regarding a potential SARS-CoV vaccine, while shedding light on mechanisms involved in SARS pathogenesis. 相似文献
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Preliminary study on the detection of the SARS-CoV specific target cDNA fragments by multiplex PCR 总被引:3,自引:0,他引:3
The multiplex polymerase chain reaction (PCR) technique was applied to detect the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) specific target cDNA fragments in the present study. The target cDNA fragments of SARS-CoV were synthesized artificially according to the genome sequence of SARS-CoV in GenBank submitted by The Chinese University of Hong Kong, and were used as simulated positive samples. Five primers recommended by World Health Organization (WHO) were used to amplify the fragments by single PCR and multiplex PCR. Three target cDNA fragments (121, 182 and 302 bp), as well as the three different combinations of any two of these fragments, were amplified by single PCR. The combination of these three fragments was amplified by multiplex PCR. The re~sults indicated that the multiplex PCR technique could be applied to detect the SARS-CoV specific target cDNA fragments successfully. 相似文献
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为了明确抗SARS-CoVN蛋白单克隆抗体的特异性,并鉴定其识别表位,首先在E.coli中表达了人类冠状病毒229E(HCoV-229E)和OC43(HCoV-OC4)N蛋白,用Westernblotting和间接免疫荧光方法分别检测了4株抗SARS-CoVN蛋白单克隆抗体(1-1C2、1-1D6、2-8F11和2-2E5)与HCoV-OC43和HCoV-229E及其N蛋白的交叉反应情况,而后应用12种重组截短型SARS-CoVN蛋白对上述4种单克隆抗体的识别表位进行了初步定位。结果显示:(1)在4株抗N蛋白单克隆抗体中,1-1C2、1-1D6和2-2E5不与HCoV-OC43和HCoV-229E及其N蛋白发生交叉反应,为SARS-CoVN蛋白特异性抗体;(2)2-8F11、1-1D6和2-2E5针对的抗原表位位于SARS-CoVN蛋白的aa30-60,1-1C2针对的抗原表位则位于SARS-CoVN蛋白的aa170-184。这一研究为阐明SARS-CoVN蛋白的免疫学特征,建立特异性免疫诊断技术和研究其致病机制提供了必要的依据和材料。 相似文献
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为探讨SARS-CoV未知蛋白在基因表达调控中的作用,本研究以鉴定的SARS-CoVBJ01毒株cDNA为材料,采用PCR方法扩增SARS-CoV未知蛋白基因Orf7,Orf8,Orf9,引入GFP和pDsRed作为报告基因,构建出5种表达质粒:pEGFP-Orf7,pEGFP-Orf8,pEGFP-Orf79,pcDNA3-Orf7,pcDNA3-Orf8。进行体外转染实验后,采用荧光酶标仪测量其细胞中荧光蛋白的表达量,再采用流式细胞仪进一步进行瞬时表达的分析比较。结果发现,pcDNA3-Orf7与pEGFP共转染时,pcDNA3-Orf7能够增强绿色荧光蛋白(GFP)的表达,pEGFP-Orf7与pDsRed共转染时pEGFP-Orf7可以增强红色荧光蛋白(RFP)的表达,且同时表达红色和绿色荧光的细胞数增加。本文首次证明了ORF7编码的63个氨基酸的蛋白具有反式激活作用,即为SARS-CoV自身的反式激活因子。 相似文献
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Bi S Qin E Xu Z Li W Wang J Hu Y Liu Y Duan S Hu J Han Y Xu J Li Y Yi Y Zhou Y Lin W Xu H Li R Zhang Z Sun H Zhu J Yu M Fan B Wu Q Lin W Tang L Yang B Li G Peng W Li W Jiang T Deng Y Liu B Shi J Deng Y Wei W Liu H Tong Z Zhang F Zhang Y Wang C Li Y Ye J Gan Y Ji J Li X Tian X Lu F Tan G Yang R Liu B Liu S Li S Wang J Wang J Cao W Yu J Dong X Yang H 《基因组蛋白质组与生物信息学报(英文版)》2003,1(3):180-192
Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the SARS-CoV.It is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving 相似文献