【摘 要】
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Yellow fever virus (YFV) is a re-emerging virus that can cause life-threatening yellow fever disease in humans.Despite the availability of an effective vaccine,little is known about the replication mechanism of YFV,and there are still no available specifi
【机 构】
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The Centre for Infection and Immunity Studies,School of Medicine,Sun Yat-Sen University,Guangzhou 51
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Yellow fever virus (YFV) is a re-emerging virus that can cause life-threatening yellow fever disease in humans.Despite the availability of an effective vaccine,little is known about the replication mechanism of YFV,and there are still no available specific anti-YFV medicines.Herein,by introducing the Renilla luciferase gene (Rluc) into an infectious clone of YFV vaccine strain 17D,we generated a recombinant virus 17D-Rluc.2A via reverse genetics approaches.The 17D-Rluc.2A had similar plaque morphology and comparable in vitro growth characteristics with its parental strain.Importantly,the reporter luciferase was efficiently expressed in 17D-Rluc.2A-infected mammalian and mosquito cells,and there was a good linear correlation between intracellular luciferase expression and extracellular infectious virion reproduction.Furthermore,by a combination of the 17D-Rluc.2A reporter virus and selective 2\'-hydroxyl acylation analyzed by primer extension (SHAPE)technology,the conserved 5\'-SLA element was shown to be essential for YFV replication,highlighting the capability of 17D-Rluc.2A in the investigation of YFV replication.At last,we demonstrated that two compounds with distinct anti-viral mechanisms can effectively inhibit the viral propagation in 17D-Rluc.2A-infected cells,demonstrating its potential application in the evaluation of anti-viral medicines.Taken together,the 17D-Rluc.2A serves as a useful tool for the study of YFV replication and anti-YFV medicine development.
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