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
Since 1995,three viral isolates named RGV9506,RGV9807 and RGV9808 associated with frog mortality were isolated from farm-raised frogs (Rana grylio virus,RGV) in China.Both ultrastructural morphology and serological ch...Since 1995,three viral isolates named RGV9506,RGV9807 and RGV9808 associated with frog mortality were isolated from farm-raised frogs (Rana grylio virus,RGV) in China.Both ultrastructural morphology and serological characterisitics had shown that the RGV isolates belong to genus Ranavirus. The DNA sequence analysis of the 5′end of the major capsid protein(MCP) gene of RGV isolates by PCR amplification produced a 531bp fragment.DNA templates were prepared from cells infected with different RGV isolates and the specific primers designed were based on highly conserved region at the 5′ of the gene encoding Frog virus 3(FV3) MCP, which is the typical species of the genus Ranavirus.The PCR products indicate that the nucleotide sequence of MCP gene of the RGV9506,RGV9807 and RGV9808 showed 99.6%, 99.8% and 98.4% homology respectively with the corresponding region of the MCP gene of FV3.展开更多
大菱鲆红体病虹彩病毒(turbot reddish body iridovirus,TRBIV)是中国工厂化养殖大菱鲆的主要病毒性病原之一。本研究提取了该病毒DNA,依据其主要衣壳蛋白基因序列设计了PCR引物,优化了PCR反应参数,建立了TRBIV的PCR检测方法,测试了该...大菱鲆红体病虹彩病毒(turbot reddish body iridovirus,TRBIV)是中国工厂化养殖大菱鲆的主要病毒性病原之一。本研究提取了该病毒DNA,依据其主要衣壳蛋白基因序列设计了PCR引物,优化了PCR反应参数,建立了TRBIV的PCR检测方法,测试了该方法的特异性和灵敏度,并应用该方法开展了TRBIV的流行情况调查及研究了大菱鲆的外观症状(红体)与TRBIV感染的关系。结果显示,本研究建立的TRBIVPCR检测方法可以从相当于100ng病鱼组织中或103数量级的病毒粒子中检出TRBIV,也可以在不杀死被测鱼的情况下,仅从鱼体中抽取少量血液即可在1天的时间内完成大量样品的TRBIV检测,但不能从健康大菱鲆脾组织和患淋巴囊肿牙鲆的囊肿组织中扩增出任何产物,说明该方法具有很高的灵敏性和特异性;在所调查的山东半岛5家大菱鲆养殖场的19尾大菱鲆中,7尾为TRBIV阳性或弱阳性;具有"红体"症状的大菱鲆应当划分为"病毒性红体病"、"细菌性红体病"和"非病原性红体症"3种不同的类型。本研究为TRBIV的流行情况和传播途径调查、疾病的快速诊断和控制提供了技术手段;调查结果显示TRBIV已遍布山东半岛沿海地区的大菱鲆主产区,在地域上相距较远的多个大菱鲆养殖场流行,今后需要密切关注该病毒的传播和流行。展开更多
基金the National Natural Science Foundation of China(No20674029) the Science and Technology Department ofJilin Province(No20050402-4)
文摘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.
文摘Since 1995,three viral isolates named RGV9506,RGV9807 and RGV9808 associated with frog mortality were isolated from farm-raised frogs (Rana grylio virus,RGV) in China.Both ultrastructural morphology and serological characterisitics had shown that the RGV isolates belong to genus Ranavirus. The DNA sequence analysis of the 5′end of the major capsid protein(MCP) gene of RGV isolates by PCR amplification produced a 531bp fragment.DNA templates were prepared from cells infected with different RGV isolates and the specific primers designed were based on highly conserved region at the 5′ of the gene encoding Frog virus 3(FV3) MCP, which is the typical species of the genus Ranavirus.The PCR products indicate that the nucleotide sequence of MCP gene of the RGV9506,RGV9807 and RGV9808 showed 99.6%, 99.8% and 98.4% homology respectively with the corresponding region of the MCP gene of FV3.
文摘大菱鲆红体病虹彩病毒(turbot reddish body iridovirus,TRBIV)是中国工厂化养殖大菱鲆的主要病毒性病原之一。本研究提取了该病毒DNA,依据其主要衣壳蛋白基因序列设计了PCR引物,优化了PCR反应参数,建立了TRBIV的PCR检测方法,测试了该方法的特异性和灵敏度,并应用该方法开展了TRBIV的流行情况调查及研究了大菱鲆的外观症状(红体)与TRBIV感染的关系。结果显示,本研究建立的TRBIVPCR检测方法可以从相当于100ng病鱼组织中或103数量级的病毒粒子中检出TRBIV,也可以在不杀死被测鱼的情况下,仅从鱼体中抽取少量血液即可在1天的时间内完成大量样品的TRBIV检测,但不能从健康大菱鲆脾组织和患淋巴囊肿牙鲆的囊肿组织中扩增出任何产物,说明该方法具有很高的灵敏性和特异性;在所调查的山东半岛5家大菱鲆养殖场的19尾大菱鲆中,7尾为TRBIV阳性或弱阳性;具有"红体"症状的大菱鲆应当划分为"病毒性红体病"、"细菌性红体病"和"非病原性红体症"3种不同的类型。本研究为TRBIV的流行情况和传播途径调查、疾病的快速诊断和控制提供了技术手段;调查结果显示TRBIV已遍布山东半岛沿海地区的大菱鲆主产区,在地域上相距较远的多个大菱鲆养殖场流行,今后需要密切关注该病毒的传播和流行。
