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Generation of duck Tembusu virus using a simple reverse genetic system in duck embryo fibroblast cells

文献类型： 外文期刊

作者： Hu, Feng 1 ; Zhu, Tong 1 ; Guo, Xiaozhen 1 ; Yu, Kexiang 1 ; Ma, Xiuli 1 ; Liu, Cunxia 1 ; Liu, Liping 1 ; Gao, Yuehua 1 ; Song, Minxun 1 ; Wu, Jiaqiang 1 ; Huang, Bing 1 ; Li, Yufeng 1 ;

作者机构： 1.Shandong Acad Agr Sci, Inst Poultry Sci, Shandong Prov Key Lab Immun & Diag Poultry Dis, 1 Jiaoxiao Rd, Jinan 250023, Shandong, Peoples R China

关键词： Duck tembusu virus; Infectious-subgenomic-amplicons; Duck embryo fibroblast cells; Infectious done

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2022 年 300 卷

页码：

收录情况： SCI

摘要： Outbreaks of duck Tembusu virus (DTMUV) have caused serious economic losses in China since 2010. In this study, an infectious clone of the DTMUV BZ-2010strain, isolated from layer cherry duck in China, was constructed using the bacterium-free infectious subgenomic-amplicons method. The subgenomic-amplicons of the human cytomegalovirus promoter (pCMV) at the 5' terminus of the first DNA fragment, the entire genome of DTMUV, and the hepatitis delta ribozyme followed by the simian virus 40 polyadenylation signal (HDR/SV40pA) at the 3' terminus of the last DNA fragment were synthesized and amplified by PCR in three DNA fragments. The pCMV and HDR/SV40pA were used to drive the viral RNA transcription and generate a full-length RNA transcript of the virus, and were found to be effective in reassembling DTMUV in duck embryo fibroblast cells. The RNA transcripts from the infection clone were infectious in duck embryo fibroblast cells, generating the reconstituted DTMUV. This study provided a valuable reverse genetic tool for the further study DTMUV pathogenesis.

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