McElroy Kate L, Tsetsarkin Konstantin A, Vanlandingham Dana L, Higgs Stephen
Department of Pathology, University of Texas Medical Branch, Keiller 2.104, 301 University Boulevard, Galveston, TX 77555-0609, USA.
J Gen Virol. 2005 Jun;86(Pt 6):1747-1751. doi: 10.1099/vir.0.80746-0.
Infectious clone technology provides an opportunity to study the molecular basis of arthropod-virus interactions in detail. This study describes the development of an infectious clone of the prototype yellow fever virus Asibi strain (YFV-As) with the purpose of identifying sequences or domains that influence infection dynamics in the mosquito vector. The full-length cDNA of YFV-As virus was produced from RT-PCR products of parental viral RNA. These were cloned into a low-copy-number plasmid previously used to develop the YFV-17D infectious clone (pACNR/FLYF-17D). Virus recovered from the infectious clone exhibited biological characteristics similar to those of the parental YFV-As, including replication kinetics, reactivity to flavivirus cross-reactive and YFV-specific antibodies and infection and dissemination rates in Aedes aegypti, the principal mosquito vector of YFV. These data provide the basis for future studies with chimeric Asibi/17D viruses to identify the determinants of vaccine attenuation in the vector.
感染性克隆技术为详细研究节肢动物与病毒相互作用的分子基础提供了契机。本研究描述了原型黄热病毒阿西比株（YFV-As）感染性克隆的构建，目的是鉴定影响蚊虫载体中感染动态的序列或结构域。YFV-As病毒的全长cDNA由亲本病毒RNA的RT-PCR产物制备而成。将这些产物克隆到先前用于构建YFV-17D感染性克隆的低拷贝数质粒（pACNR/FLYF-17D）中。从感染性克隆中获得的病毒表现出与亲本YFV-As相似的生物学特性，包括复制动力学、对黄病毒交叉反应性抗体和YFV特异性抗体的反应性以及在黄热病毒主要蚊虫载体埃及伊蚊中的感染和传播率。这些数据为未来利用嵌合阿西比/17D病毒进行研究以确定载体中疫苗减毒的决定因素奠定了基础。
