目的研究狂犬病毒糖蛋白基因重排至基因组的第二位对病毒生物学特性的影响。方法应用基因操作技术,将狂犬病毒HEP-Flury株全基因组中G基因从原来的第四位重排至第二位,构建重组病毒N1G2的全长感染性cDNA克隆,拯救重组病毒N1G2。再将重组病毒与亲代毒株比较,分析两者致病性、免疫原性及在细胞中繁殖效率的差异。结果重组病毒在BHK-21细胞中连传5代,病毒滴度开始呈现恒定,至第6-10代,病毒滴度达到10～6FFU/mL。比较重组病毒N1G2与亲代病毒rHEP-Flury株生物学特性发现:两株病毒在BHK-21细胞中增殖规律均一致,滴度为10～6FFU/mL的病毒。以MOI=0.01感染BHK-21细胞,4天后病毒滴度达到最高。用荧光定量PCR方法比较两株病毒在BHK-21细胞中G蛋白mRNA表达量,重组病毒N1G2 G mRNA表达量是亲代病毒的1.58倍。薄层扫描显示,重组病毒N1G2 G蛋白表达量高于rHEP-Flury株。动物试验表明,两个毒株对成年鼠均无致死性,与rHEP-Flury株相比,重组病毒对小鼠致病性降低,免疫原性提高。结论上述实验结果表明,狂犬病毒G基因重排至第二位,其糖蛋白表达量增高,病毒毒力减弱,病毒在BHK21细胞中捣乱繁殖效率差异不大。
Nelson Bay orthoreovirus(NBV) is a species within the genus Orthoreovirus,family Reoviridae,containing 4, possibly 5,*** we report a putative sixth member,Xi River virus(XRV),isolated from fruit bats collected in a lo...
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Nelson Bay orthoreovirus(NBV) is a species within the genus Orthoreovirus,family Reoviridae,containing 4, possibly 5,*** we report a putative sixth member,Xi River virus(XRV),isolated from fruit bats collected in a location near the Xi River,Guangdong Province,*** virus showed the same electron microscopic morphology as NBV,fusogenic CPE and a 10 fragment double-strand RNA genome,as well as high sequence identity to NBV *** is the first bat reovirus isolated in China.
As one of the oldest diseases in the world,rabies history in China can be traced back to 2000 years ago,and for more than a decade,China has been a country with the second highest human rabies cases in the *** studies...
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As one of the oldest diseases in the world,rabies history in China can be traced back to 2000 years ago,and for more than a decade,China has been a country with the second highest human rabies cases in the *** studies have revealed a profound understanding about the biodiversity of rabies virus in China,but the evolutionary dynamics of this virus in the country remain *** present study,the complete G gene sequences of 33 RABVs isolated from distinct provinces of China were determined and phylogenetic analysis was conducted using these G sequences and 91 others retrieved from GenBank representing China and Southeast *** evolutionary and divergence times of RABV were estimated using a Bayesian Markov chain Monte Carlo method to understand the temporal and spatial dynamics of this virus in China and *** showed that rabies viruses in China and Southeast Asia share a common ancestor and formed 2 clades,Asian 1 and Asian 2,with each being further divided into 3 *** time of the most recent common ancestor of current RABV variants in China and Southeast Asia was estimated to be year 1762(1648-1859) and the viruses circulating in Southeast Asia likely derived from *** study presents the first temporal-spatial dynamics of RABV in China and Southeast Asia.
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