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大口黑鲈蛙虹彩病毒MCP蛋白多克隆抗体的制备与应用
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作者 王若瑄 罗霞 +5 位作者 李宁求 林强 牛银杰 梁红茹 付小哲 吕爱军 《西北农林科技大学学报(自然科学版)》 CSCD 北大核心 2024年第4期21-27,共7页
【目的】制备大口黑鲈蛙虹彩病毒主衣壳(MCP)蛋白兔多克隆抗体,以期为该病毒蛋白功能研究奠定基础。【方法】对pET32a(+)MCP/BL21重组大肠杆菌进行诱导表达,将重组蛋白进行纯化、复性,以此为抗原免疫大耳兔制备多克隆抗体,ELISA检测抗... 【目的】制备大口黑鲈蛙虹彩病毒主衣壳(MCP)蛋白兔多克隆抗体,以期为该病毒蛋白功能研究奠定基础。【方法】对pET32a(+)MCP/BL21重组大肠杆菌进行诱导表达,将重组蛋白进行纯化、复性,以此为抗原免疫大耳兔制备多克隆抗体,ELISA检测抗体效价,间接免疫荧光试验(IFA)和Western Blot法分析MCP多克隆抗体的特异性。【结果】纯化的MCP重组蛋白条带特异;间接ELISA结果显示,制备的兔多克隆抗体血清效价为1∶1024000;IFA和Western Blot检测结果表明该多抗特异性良好,能够与大口黑鲈蛙虹彩病毒MCP蛋白发生特异性反应,IFA试验表明血清最适稀释度为1∶500。【结论】成功制备了大口黑鲈蛙虹彩病毒MCP兔多克隆抗体,该抗体可特异性识别大口黑鲈蛙虹彩病毒MCP蛋白。 展开更多
关键词 大口黑鲈 蛙虹彩病毒 主衣壳蛋白 多克隆抗体
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黄柏碱调节MCP-1/CCR2信号通路对特应性皮炎大鼠炎症反应的影响
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作者 于现朝 寇旺 杨洋 《中国美容医学》 CAS 2024年第10期13-17,共5页
目的:探讨黄柏碱对特应性皮炎(Atopic dermatitis,AD)大鼠炎症反应的影响及其作用机制。方法:构建AD大鼠模型,大鼠分为正常组、模型组、黄柏碱低剂量组、黄柏碱高剂量组、黄柏碱+巨噬细胞趋化蛋白-1(Monocyte chemotactic protein-1,MCP... 目的:探讨黄柏碱对特应性皮炎(Atopic dermatitis,AD)大鼠炎症反应的影响及其作用机制。方法:构建AD大鼠模型,大鼠分为正常组、模型组、黄柏碱低剂量组、黄柏碱高剂量组、黄柏碱+巨噬细胞趋化蛋白-1(Monocyte chemotactic protein-1,MCP-1)组;给药结束后,各组大鼠做皮损评分,记录搔抓次数,HE染色观察皮肤组织病理变化,甲苯胺蓝染色测定肥大细胞数,ELISA试剂盒测定皮肤组织活性氧(Reactiveoxygenspecies,ROS)和血清免疫球蛋白(Immunoglobulins,IgE)、白介素-17(Interleukin,IL-17)、IL-6的含量,Western blot检测皮肤组织MCP-1、CC类趋化因子受体2(CC chemokinereceptor 2,CCR2)蛋白表达。结果:与正常组相比,模型组大鼠脱毛处皮肤发生溃疡、红斑,表皮和棘层增厚且有大量炎性细胞浸润,搔抓次数、肥大细胞数、ROS、IgE、IL-17、IL-6及MCP-1、CCR2蛋白表达水平增加(P<0.05);与模型组相比,黄柏碱低、高剂量组皮肤组织红斑、溃疡情况、炎性细胞浸润减少,未见脱屑,搔抓次数、肥大细胞数、ROS、IgE、IL-17、IL-6及MCP-1、CCR2蛋白表达水平依次降低(P<0.05);与黄柏碱高剂量组相比,黄柏碱+MCP-1组皮肤红斑、溃疡和炎性细胞浸润加重,搔抓次数、肥大细胞数、ROS、IgE、IL-17、IL-6及MCP-1、CCR2蛋白表达水平增加(P<0.05)。结论:黄柏碱可能通过抑制MCP-1/CCR2信号通路降低AD大鼠炎症反应。 展开更多
关键词 特应性皮炎 炎症反应 黄柏碱 mcp-1/CCR2信号通路 巨噬细胞趋化蛋白-1
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Assembly and Immunogenicity of Human Papillomavirus Type 16 Major Capsid Protein(HPV16 L1) in Pichia pastoris
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作者 LIU Da-wei ZHANG Yu YU Xiang-hui JIANG Chun-lai CHEN Yue WU Yong-ge JIN Ying-hua NIU Jun Qu Ning LIU Ming KONG Wei 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2007年第2期200-203,共4页
In this study, a recombinant Pichia pastoris expression system was developed to express HPV16 L1 protein that was driven by a strong AOX1 promoter. HPV16L1 gene was cloned into vector pPICZ,αB. HPV16 L1 protein expre... In this study, a recombinant Pichia pastoris expression system was developed to express HPV16 L1 protein that was driven by a strong AOX1 promoter. HPV16L1 gene was cloned into vector pPICZ,αB. HPV16 L1 protein expression induced by methanol was screened by using sodium dedecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE) and Western blotting. The results indicate that the HPVl6 L1 protein is secreted by the recombinant P. pastoris, and the purified HPV16 L1 protein can self-assemble into vires-like particles( VLPs), which show a good immunogenicity and induces high-titer antibody in mice. 展开更多
关键词 Human papillomavirus major capsid protein Recombinant Pichia pastoris Vires-like particles
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血清MCP-1、TRAF-6水平在脓毒症严重程度和急性肾损伤评估的作用 被引量:3
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作者 郭峻氚 陈东 +2 位作者 郭仁楠 肖东 刘艳 《新疆医科大学学报》 CAS 2023年第4期485-489,共5页
目的探讨血清单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子受体相关因子-6(TRAF-6)水平在脓毒症严重程度和急性肾损伤评估中的作用。方法回顾性分析2021年6月至2022年6月在新疆维吾尔自治区人民医院重症医学科接受治疗的110例脓毒症患者的... 目的探讨血清单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子受体相关因子-6(TRAF-6)水平在脓毒症严重程度和急性肾损伤评估中的作用。方法回顾性分析2021年6月至2022年6月在新疆维吾尔自治区人民医院重症医学科接受治疗的110例脓毒症患者的病例资料,依据脓毒症相关急性肾损伤发生情况分为发生组(50例)、未发生组(60例)。统计分析两组患者基线资料、血清MCP-1、TRAF-6水平,并分析脓毒症患者血清MCP-1、TRAF-6水平与序贯多器官功能障碍(SOFA)评分、急性生理和慢性健康Ⅱ(APACHEⅡ)评分的相关性,多因素Logistic回归分析脓毒症相关急性肾损伤影响因素。结果110例患者中,脓毒症相关急性肾损伤发生50例,未发生60例,发生率为45.45%。发生组患者的高血压、肺部感染比例均高于未发生组(P<0.05),SOFA评分、APACHEⅡ评分均高于未发生组(P<0.05),氧合指数低于未发生组(P<0.05),肾小球滤过率(eGFR)、血肌酐(SCr)、尿素氮(BUN)水平均高于未发生组(P<0.05),动脉血乳酸水平高于未发生组(P<0.05),但两组患者的性别、年龄、糖尿病比例的差异无统计学意义(P>0.05)。发生组患者的血清MCP-1、TRAF-6水平均高于未发生组(P<0.05)。脓毒症患者血清MCP-1、TRAF-6水平与SOFA评分、APACHEⅡ评分均呈显著的正相关关系(P<0.05)。多因素Logistic回归分析显示,脓毒症相关急性肾损伤影响因素包括高血压、肺部感染、SOFA评分、APACHEⅡ评分、肾小球滤过率(eGFR)、动脉血乳酸、MCP-1、TRAF-6(P<0.05),不包括氧合指数、尿素氮(BUN)、SCr(P>0.05)。结论血清MCP-1、TRAF-6水平和脓毒症严重程度和关系密切,可作为急性肾损伤的诊断参考指标。 展开更多
关键词 脓毒症 严重程度 急性肾损伤 mcp-1 TRAF-6
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Bioinformatic Analysis of Non-VP1 Capsid Protein of Coxsackievirus A6 被引量:4
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作者 刘洪波 阳广菲 +1 位作者 梁思佳 林军 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2016年第4期607-613,共7页
This study bioinformatically analyzed the non-VP1 capsid proteins(VP2-VP4) of Coxasckievirus A6(CVA6), with an attempt to predict their basic physicochemical properties, structural/functional features and linear B... This study bioinformatically analyzed the non-VP1 capsid proteins(VP2-VP4) of Coxasckievirus A6(CVA6), with an attempt to predict their basic physicochemical properties, structural/functional features and linear B cell eiptopes. The online tools Sub Loc, Target P and the others from Ex PASy Bioinformatics Resource Portal, and SWISS-MODEL(an online protein structure modeling server), were utilized to analyze the amino acid(AA) sequences of VP2-VP4 proteins of CVA6. Our results showed that the VP proteins of CVA6 were all of hydrophilic nature, contained phosphorylation and glycosylation sites and harbored no signal peptide sequences and acetylation sites. Except VP3, the other proteins did not have transmembrane helix structure and nuclear localization signal sequences. Random coils were the major conformation of the secondary structure of the capsid proteins. Analysis of the linear B cell epitopes by employing Bepipred showed that the average antigenic indices(AI) of individual VP proteins were all greater than 0 and the average AI of VP4 was substantially higher than that of VP2 and VP3. The VP proteins all contained a number of potential B cell epitopes and some eiptopes were located at the internal side of the viral capsid or were buried. We successfully predicted the fundamental physicochemical properties, structural/functional features and the linear B cell eiptopes and found that different VP proteins share some common features and each has its unique attributes. These findings will help us understand the pathogenicity of CVA6 and develop related vaccines and immunodiagnostic reagents. 展开更多
关键词 Coxsackievirus A6 (CVA6) capsid proteins bioinformatics physicochemical properties structural and functional domains linear B cell eiptopes
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Expression and Characterization of a Recombinant Truncated Capsid Protein of Hepatitis E Virus in Pichia pastoris 被引量:2
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作者 YANG En-cheng CHI Bao-rong +7 位作者 LI Xiao LIU Yan GAO Peng JIA Peng KAN Shi-fu WEN Zhong-mei WANG Wan JIN Ning-yi 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2010年第2期235-239,共5页
Hepatitis E is an enterically transmitted viral disease caused by infection with hepatitis E virus(HEV). HEV is a nonenveloped virus that bas been classified in the family of Caliciviridae. The virus appears to be a... Hepatitis E is an enterically transmitted viral disease caused by infection with hepatitis E virus(HEV). HEV is a nonenveloped virus that bas been classified in the family of Caliciviridae. The virus appears to be a polya-denylated, positive-stranded RNA virus with three major open reading frames(ORFs). The capsid protein of HEV is encoded by the open reading frame 2(ORF2). We attempted to produce a truncated capsid protein, designed p293, in Pichia pastoris. The p293 gene encoding amino acids(aa) 382-674 of HEV ORF2 was designed based on the full length of HEV ORF2, cloned into the yeast vector pPIC9K, and expressed in P. pastoris strain GS 115. SDS-PAGE and Western blotting demonstrated that the recombinant protein p293 could well be expressed in P pastoris. Under optimized conditions (culture medium pH, 6.0-6.5; methanol concentration added daily, 3.0%; inoculum density, OD600=60; induction time point, 72-96 h), the yield of soluble p293 was approximately 80 mg/L. We also observed p293 secretory expressed in P. pastoris to be 30 nm viral like particles by using electron microscopy. These results show that the p293 may has utility in the analysis of cell specific factors in the protein processing and assembly of HEV, and serve as a useful antigen for both diagnostic and vaccine purposes. 展开更多
关键词 Hepatitis E virus capsid protein PICHIAPASTORIS protein purification
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Prokaryotic Expression and Potential Application of the Truncated PCV-2 Capsid Protein 被引量:4
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作者 Zhong-zi LOU Zhi-yong LI +6 位作者 Gang WANG Jian-qiang LI Xi LAN Xue-rui LI Xiang-ping YIN Ji-xing LIU Si-dang LIU 《Virologica Sinica》 SCIE CAS CSCD 2010年第2期86-97,共12页
Three pairs of specific primers were designed to amplify the F2-1, F2-2 and XF2-2 truncated sequences of ORF2 which encodes the capsid protein of porcine circovirus type 2 (PCV-2). The F2-1 sequence had most of the ... Three pairs of specific primers were designed to amplify the F2-1, F2-2 and XF2-2 truncated sequences of ORF2 which encodes the capsid protein of porcine circovirus type 2 (PCV-2). The F2-1 sequence had most of the NLS region of ORF2, but the F2-2 and XF2-2 genes had the NLS region deleted. Truncated genes were subcloned into pET-32a(+) vectors to construct recombinant fusion expression vectors. The vectors were then transformed into Rosetta(DE3) E. coli and expressed by induction of IPTG. Expressed proteins were detected by western blotting and ELISA. The protein with best immunoreactivity was confirmed and selected, then utilized to inoculate SPF rabbits to prepare polyclonal antibodies. The protein and prepared polyclonal antibody were utilized to detect sera samples against PCV-2 from Shandong province and PCV-2 particles in PK-15 cells. In our study, three recombinant fusion proteins were successfully obtained, and the molecular weights of fusion proteins were 35.9 kDa, 33.6 kDa and 38.6 kDa respectively detected by SDS-PAGE. All of the proteins showed positive reaction with anti-PCV-2 antisera, and His-XF2-2 showed better immunoreactivity than the others. The protein of His-XF2-2 was coated as antigen in ELISA to detect the seroprevalence of PCV-2 in certain districts of Shandong province, the seropositivity rate was 27.7 % (73/264). Specific fluorescence and positive signals for PCV-2 could be detected in PK-15 cells inoculated with PCV-2 with the participation of prepared antibodies against His-XF2-2 in IFA and IPMA. Experimental results indicated that the truncated PCV-20RF2 gene containing most of the NLS region was successfully expressed in E. coli, and His-XF2-2 was demonstrated to have better immunoreactivity with anti-PCV-2 antisera than the other two fusion proteins. His-XF2-2 and prepared polyclonal antibodies against it had a satisfactory capability in detecting PCV-2 infection. 展开更多
关键词 Porcine circovirus type 2 capsid protein Fusion expression Polyclonal antibodies Virus detection
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Expression of Outer Capsid Protein VP5 of Grass Carp Reovirus in E.coli and Analysis of its Immunogenicity 被引量:5
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作者 Lan-lan ZHANG Jin-yu SHEN +3 位作者 Cheng-feng LEI Chao FAN Gui-jie HAO Qin FANG 《Virologica Sinica》 SCIE CAS CSCD 2009年第6期545-551,共7页
Grass carp reovirus (GCRV) is a tentative member of the Aquareovirus genus in the family Reoviridae. The mature virion comprises 11 dsRNA genomes enclosed by two concentric icosahedral proteins shells that is comprise... Grass carp reovirus (GCRV) is a tentative member of the Aquareovirus genus in the family Reoviridae. The mature virion comprises 11 dsRNA genomes enclosed by two concentric icosahedral proteins shells that is comprised of five core proteins and two outer capsid proteins. The genome sequence and 3D structure demonstrate there is a higher level of sequence homology in structural proteins between GCRV and mammalian orthoreoviruses (MRV) compared to other members of the family. To understand the pathogenesis of GCRV infection, the outer capsid protein VP5, a homology of the μ1 protein of MRV, was expressed in E.coli. It was found that the recombinant VP5 was highly expressed, and the expressed His-tag fusion protein was involved in the formation of the inclusion body. Additionally, specific anti-VP5 serum was prepared from purified protein and western blot demonstrated that the expressed protein was able to bind immunologically to rabbit anti GCRV particle serum and the immunogenicity was determined by ELISA assay. Additional experiments in investigating the functional properties of VP5 will further elucidate the role of the GCRV outer capsid protein VP5 during entry into host cells, and its interaction among viral proteins and host cells during the infection process. 展开更多
关键词 Grass carp reovirus (GCRV) Outer capsid protein VP5 Expression in E.coli IMMUNOGENICITY
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Expression and Immunological Analysis of Capsid Protein Precursor of Swine Vesicular Disease Virus HK/70 被引量:3
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作者 Hong TIAN Jing-yan WU You-jun SHANG Shuang-hui YING Hai-xue ZHENG Xiang-tao LIU 《Virologica Sinica》 SCIE CAS CSCD 2010年第3期206-212,共7页
VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability... VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore, anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection. 展开更多
关键词 Swine vesicular disease virus capsid protein precursor gene (vp1) Gene expression Immunere sponse
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鸭坦布苏病毒Capsid蛋白原核表达及多克隆抗体制备
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作者 焦琳琳 成玉婷 +4 位作者 吴庆国 吴双 吴植 朱善元 钱莺娟 《中国畜牧兽医》 CAS CSCD 北大核心 2023年第6期2395-2402,共8页
【目的】本研究选择原核表达系统表达鸭坦布苏病毒(Duck Tembusu virus,DTMUV)核衣壳蛋白(Capsid protein),并制备其多克隆抗体,为DTMUV分子机制研究奠定基础。【方法】根据DTMUV-201909株基因序列,运用一步克隆技术将Capsid基因克隆至... 【目的】本研究选择原核表达系统表达鸭坦布苏病毒(Duck Tembusu virus,DTMUV)核衣壳蛋白(Capsid protein),并制备其多克隆抗体,为DTMUV分子机制研究奠定基础。【方法】根据DTMUV-201909株基因序列,运用一步克隆技术将Capsid基因克隆至表达载体pET-30a(+)中,将重组质粒转化大肠杆菌BL21(DE3)感受态细胞,利用IPTG进行诱导表达,通过SDS-PAGE和Western blotting鉴定重组蛋白;使用ISA206佐剂与纯化后的重组蛋白混合乳化后免疫BALB/c小鼠,以获得多克隆抗体。间接ELISA方法测定获得的多克隆抗体效价,并对多克隆抗体进行Western blotting和间接免疫荧光试验(IFA)验证。【结果】试验成功构建pET-30a-Capsid重组质粒,SDS-PAGE结果显示,表达的重组蛋白大小约为18 ku,主要以包涵体形式存在;Western blotting结果表明,该蛋白能与抗His标签鼠单克隆抗体发生特异性反应,具有良好反应原性。间接ELISA结果显示,制备的鼠抗Capisd蛋白多克隆抗体效价可达1∶256000;Western blotting和IFA结果显示,制备的多克隆抗体能特异性识别DTMUV感染细胞样品的Capsid蛋白。【结论】本研究成功制备小鼠抗Capsid蛋白多克隆抗体,为深入研究DTMUV Capsid蛋白的结构和功能提供了试验材料,为进一步阐明DTMUV的致病机制奠定基础。 展开更多
关键词 鸭坦布苏病毒(DTMUV) 核衣壳蛋白 原核表达 多克隆抗体
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Expression of the Capsid Precursor Protein gene of Foot-and-mouth Disease Virus and Green Fluorescent Protein Gene in BHK-21 Cells Mediated by Retroviral Vector
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作者 LI Jiong LIU Yan-hong +4 位作者 AN Fang-lan LIU Jun-lin LIU Xiang-tao SHANG You-jun YIN Hong 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期70-75,共6页
We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constr... We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine. 展开更多
关键词 retroviral vector FMDV capsid precursor protein gene green fluorescent protein gene BHK-21 cell
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Homology Modelling and Structural Comparisons of Capsid-Associated Proteins from Circoviruses Reveal Important Virus-Specific Surface Antigens 被引量:1
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作者 Edward I. Patterson Jade K. Forwood Shane R. Raidal 《Crystal Structure Theory and Applications》 2012年第2期9-16,共8页
Circoviridae represent a growing family of small animal viruses. Some of these viruses have veterinary and medical importance, although, a vast amount of these newly discovered viruses have unknown effects on their ho... Circoviridae represent a growing family of small animal viruses. Some of these viruses have veterinary and medical importance, although, a vast amount of these newly discovered viruses have unknown effects on their hosts. The capsid-associated protein (Cap) of circoviruses is of interest because of its role in viral structure, immune evasion, host cell entry, and nuclear shuttling of viral components. The structure of the porcine circovirus 2 (PCV2) Cap has been solved and offered insight to these functions. Based on the crystallographic PCV2 Cap structure, models from circoviruses isolated from avian, fish, and mammalian hosts have been constructed and analyzed to better understand the roles of these proteins in the virus family. A high degree of conservation is observed in the models, however, the surface antigens differ among viruses. This is likely a reflection of the small genome harbored by circoviruses, and therefore the requirement of their few proteins to carry out specific vital functions, while maintaining enough variation to successfully infect their hosts. Here we describe the putative structures of a range of Cap proteins from circoviruses based on the crystallographic determination of porcine Cap, identifying key regions for function and inhibition of crystal formation. 展开更多
关键词 CIRCOVIRUS capsid-Associated protein Structure HOMOLOGY CAP Modelling
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A Novel Pharmacophore Model Derived from a Class of Capsid Protein Enterovirus 71 Inhibitors
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作者 段红霞 杨新玲 +3 位作者 王道全 宁君 梅向东 张健 《Chinese Journal of Structural Chemistry》 SCIE CAS CSCD 2012年第8期1159-1169,共11页
Capsid protein enterovirus 71 (EV71) is one of the major viruses that cause the severe encephalitis and thus result in a high mortality in children less than 5 years of age.In an effort to discover new potent inhibi... Capsid protein enterovirus 71 (EV71) is one of the major viruses that cause the severe encephalitis and thus result in a high mortality in children less than 5 years of age.In an effort to discover new potent inhibitors against EV71,a novel three-dimensional pharmacophore model was developed on 24 inhibitors with different molecular structures and bioactivities.The best hypothesis (Hypo1) has a high predictive power and consists of four features,namely,one hydrophobic point (HY) and three hydrogen-bond acceptors (HA).Two key features of the best Hypo1,HY1 and HA3 match well with an important narrow hydrophobic canyon and with the surface of LYS274 in the target EV71 active site,respectively.The more versatile feature,HA1,is firstly found to be very influential on these compounds’ bioactivities,which may interact with the other side of the active site in the EV71 receptor.The application of the model is successful in predicting the activities of 30 known EV71 inhibitors with a correlation coefficient of 0.831.Furthermore,Hypo1 demonstrates a superior screening capability for retrieving inhibitors from the database with a high enrichment factor of 70.This study provides some important clues in search for more potent inhibitors against EV71 infection. 展开更多
关键词 capsid protein enterovirus 71 inhibitor hand-foot-and-mouth disease pharmacophore model hydrogen-bond acceptor hydrophobic point
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Interactions of the HSV-1 UL25 Capsid Protein with Cellular Microtubule-associated Protein
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作者 Lei GUO Ying ZHANG Yan-chun CHE Wen-juan WU Wei-zhong LI Li-chun WANG Yun LIAO Long-ding LIU Qi-han LI 《Virologica Sinica》 SCIE CAS CSCD 2008年第3期211-217,共7页
An interaction between the HSV-1 UL25 capsid protein and cellular microtubule-associated protein was found using a yeast two-hybrid screen and β-D-galactosidase activity assays. Immunofluorescence microscopy of the U... An interaction between the HSV-1 UL25 capsid protein and cellular microtubule-associated protein was found using a yeast two-hybrid screen and β-D-galactosidase activity assays. Immunofluorescence microscopy of the UL25 protein demonstrated its co-localization with cellular microtubule-associated protein in the plasma membrane. Further investigations with deletion mutants suggest that UL25 is likely to have a function in the nucleus. 展开更多
关键词 HSV-1 capsid UL25 Microtubule-associated protein
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Research Progress of HPV L1 Capsid Protein in Prediction of Cervical Lesions
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作者 Jing Cheng Lin Xu +5 位作者 Beibei Liu Biao Wang Xicui Long Zhihong Li Ruiting Wu Ruili Chen 《Proceedings of Anticancer Research》 2020年第6期36-40,共5页
Cervical cancer is one of the most common malignant gynecological tumors and has the second highest incidence of all malignancies in females.Chronic and persistent infection with High Risk Human Papillomavirus(HR-HPV)... Cervical cancer is one of the most common malignant gynecological tumors and has the second highest incidence of all malignancies in females.Chronic and persistent infection with High Risk Human Papillomavirus(HR-HPV)is the main cause of cervical cancer.There is a distinct lack of methodology by which to determine whether cervical epithelial dysplasia is cancerous following HPV infection.HPV L1 capsid protein is a major structural protein of human papillomavirus(HPV),and it is the main target of the local cellular immune response aiming to combat human papillomavirus after HPV infection within cervical cells.Greater understanding of HPV L1 capsid protein and its association with cervical cytology,histopathology,patient age and human papillomavirus viral load has the potential to contribute toward improved the diagnosis and management of cervical cancer,providing useful information for gynecological clinicians in the hope of improving patient treatment and quality of life.This article reviews the predictive utility of HPV L1 capsid protein for cervical lesions. 展开更多
关键词 HPV L1 capsid protein Cervical lesions PROGNOSIS
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Development of an indirect immunofluorescence assay for PCV3 antibody detection based on capsid protein
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作者 Lun Yao Chang Li +7 位作者 Junwei Wang Yufang Cheng Ahmed H.Ghonaim Qi Sun Xuexiang Yu Weijie Niu Shengxian Fan Qigai He 《Animal Diseases》 2021年第2期125-132,共8页
Porcine circovirus type 3(PCV3)is a novel porcine circovirus associated with porcine dermatitis and nephritis syndrome(PDNS),reproductive failure,and multisystemic inflammation.Capsid protein(Cap)encoded by PCV3 ORF2 ... Porcine circovirus type 3(PCV3)is a novel porcine circovirus associated with porcine dermatitis and nephritis syndrome(PDNS),reproductive failure,and multisystemic inflammation.Capsid protein(Cap)encoded by PCV3 ORF2 gene has been identified as an immunogenic protein.Currently,there is no immunofluorescence assay(IFA)available for serological diagnosis.Here,the N-terminal 33 amino acids of Cap protein were predicted to serve as a PCV3 nuclear localization signal(NLS).Two types of recombinant plasmids were constructed for recombinant protein expression in 5f9 cells by using a baculovirus expression system:plasmid rvBac-Pc for full-length Cap protein expression and rvBac-Sc for Cap protein expression with a honeybee melittin signal peptide in place of the predicted NLS sequence.Expression of the nuclear localization sequences was further analyzed by IFA.Strong and specific fluorescence signals were observed in the nucleus of rvBac-Pc-transfected cells and in the cytoplasm of rvBac-Sc-transfected cells.No cross-reactivity was observed with porcine circovirus type 2,porcine pseudorabies virus,classical swine fever virus,or porcine reproductive and respiratory syndrome virus.In summary,we developed two fluorescence detection modes for Cap protein that can be used to detect PCV3 antibodies.This method is suitable for the diagnosis and epidemiological investigation of PCV3.This study provides a reliable detection method for monitoring PCV3 antibody level in pigs in the future. 展开更多
关键词 PCV3 capsid protein ANTIBODIES IFA
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大口黑鲈溃疡综合征病毒MCP基因序列分析及PCR快速检测方法的建立 被引量:10
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作者 马冬梅 白俊杰 +3 位作者 邓国成 李胜杰 叶星 江小燕 《中国水产科学》 CAS CSCD 北大核心 2010年第6期1149-1156,共8页
为了早期快速诊断近年来流行于广东省养殖大口黑鲈(Micropterussalmoides)中的病毒性溃疡综合征,本研究用基因组步移的方法获得了大口黑鲈溃疡综合征病毒(Largemouthbassulcerativesyndromevirus,LBUSV)主要衣壳蛋白(MCP)基因,该基因编... 为了早期快速诊断近年来流行于广东省养殖大口黑鲈(Micropterussalmoides)中的病毒性溃疡综合征,本研究用基因组步移的方法获得了大口黑鲈溃疡综合征病毒(Largemouthbassulcerativesyndromevirus,LBUSV)主要衣壳蛋白(MCP)基因,该基因编码区全长1392bp。通过序列比较分析,在MCP基因内确定了一段241bp的特异性较强的片段作为靶序列,设计并合成引物,经过优化PCR反应条件,建立了可以快速检测大口黑鲈溃疡综合征病毒的PCR方法。实验表明,在PCR进行到30个循环反应时可以检测到的质粒最小浓度是104拷贝数/μL,相当于104个病毒粒子。利用该方法,从天然感染LBUSV的大口黑鲈脾脏组织DNA可扩增出241bp的片段,而健康大口黑鲈和感染了传染性脾肾坏死病毒样病毒的大口黑鲈脾脏组织则没有扩增条带。本研究建立的PCR检测方法具有检测快速、成本低、准确性高的特点,适用于大范围早期病害诊断的推广应用。 展开更多
关键词 大口黑鲈溃疡综合征病毒 大口黑鲈 虹彩病毒 主要衣壳蛋白 序列分析 PCR检测
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1-MCP对苹果果实贮藏期间乙烯合成代谢的影响 被引量:45
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作者 李富军 翟衡 +3 位作者 杨洪强 张新华 束怀瑞 周杰 《中国农业科学》 CAS CSCD 北大核心 2004年第5期734-738,共5页
以红富士(MalusdomesticaBorkhvar.RedFuji)苹果果实为试材,研究了1-MCP(1-methylcyclopropene,1-甲基环丙烯)对果实贮藏期间乙烯生物合成代谢过程的影响。结果表明,1-MCP维持果实硬度、降低果实的呼吸速率、脂氧合酶活性和乙烯的释放... 以红富士(MalusdomesticaBorkhvar.RedFuji)苹果果实为试材,研究了1-MCP(1-methylcyclopropene,1-甲基环丙烯)对果实贮藏期间乙烯生物合成代谢过程的影响。结果表明,1-MCP维持果实硬度、降低果实的呼吸速率、脂氧合酶活性和乙烯的释放。进一步研究表明,1-MCP处理抑制果实贮藏15d后体内ACS活性,增加ACC含量的积累,延迟ACO活性高峰的出现,并抑制果实乙烯跃变期间蛋白激酶活性的升高。 展开更多
关键词 苹果 果实贮藏期 乙烯 合成代谢 1-mcp 1-甲基环丙烯 蛋白激酶活性
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嵌杯病毒主要衣壳蛋白的研究进展
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作者 谈晓梅 徐彦召 +1 位作者 刘光清 孟春春 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第8期3354-3361,共8页
嵌杯病毒(calicivirus)是无囊膜的单股正链RNA病毒,具有广泛的宿主范围因而严重威胁着人和许多其他脊椎动物的健康。由于该病毒种属流行株多变、极易传播且呈现区域性感染,致使全球范围内因杯状病毒引起的公共卫生疾病频繁暴发,近年来,... 嵌杯病毒(calicivirus)是无囊膜的单股正链RNA病毒,具有广泛的宿主范围因而严重威胁着人和许多其他脊椎动物的健康。由于该病毒种属流行株多变、极易传播且呈现区域性感染,致使全球范围内因杯状病毒引起的公共卫生疾病频繁暴发,近年来,其感染发病率呈明显增加的趋势。不同属成员虽然感染不同种类的宿主,但其主要衣壳蛋白的结构类似。嵌杯病毒主要衣壳蛋白中含有免疫原性关键区域,能与宿主受体结合并可能介导感染过程,且氨基酸相关的免疫原性及受体结合位点的变异是公认的病毒进化的驱动力。本文就嵌杯病毒主要衣壳蛋白的基因组结构、免疫原性、受体结合情况以及遗传演化规律等方面的研究进展进行综述,以期为后续科学研究以及该病的有效防控提供参考。 展开更多
关键词 嵌杯病毒 主要衣壳蛋白 结构 功能
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‘嘎拉’苹果对不同浓度1-MCP处理的反应 被引量:30
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作者 王瑞庆 马书尚 +2 位作者 武春林 王赵改 唐燕 《西北植物学报》 CAS CSCD 北大核心 2005年第2期256-261,共6页
以 嘎拉 Kid'sOrange × Delicious' 苹果为试材,研究了0℃贮藏期间及贮藏30、60、90和120d后转入室温7d货架期间1-MCP 1-甲基环丙烯 对果实呼吸速率、乙烯释放速率、品质和蛋白质变化的影响.结果表明,1-MCP处理显著抑制... 以 嘎拉 Kid'sOrange × Delicious' 苹果为试材,研究了0℃贮藏期间及贮藏30、60、90和120d后转入室温7d货架期间1-MCP 1-甲基环丙烯 对果实呼吸速率、乙烯释放速率、品质和蛋白质变化的影响.结果表明,1-MCP处理显著抑制嘎拉苹果贮藏期间和贮后货架期间呼吸和乙烯释放速率,延缓果实硬度、可滴定酸含量的下降,对可溶性固形物含量无影响.对照果实在贮藏过程中,出现5条明显的特异性蛋白条带,1-MCP处理能抑制特异蛋白表达.300nL·L-1浓度1-MCP处理与600nL·L-11-MCP处理作用效果无显著差异. 展开更多
关键词 苹果 1-mcp 呼吸 乙烯 品质 特异蛋白
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