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Ebola virus nucleocapsid-like particles demonstrate consistent assembly, but a wide range of helical condensation

Nature Communications. 2016; 
Maofeng Jing, Baodian Guo, Haiyang Li, Bo Yang, Haonan Wang, Guanghui Kong, Yao Zhao, Huawei Xu, Yan Wang, Wenwu Ye, Suomeng Dong, Yongli Qiao, Brett M. Tyler, Wenbo Ma & Yuanchao Wang
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Gene Synthesis Plasmids encoding Ebola virus structural proteins NP, VP35 and VP24 were originally received from Dr. Gary Nabel (NIH/VRC) in the plasmid pVR2000; however, due to expression difficulties, new plasmids were designed (Huang et al. 2002). cDNA encoding Ebola virus NP, VP24, VP35, VP30, VP40, and GP (GenBank Accession # EU224440.2) were individually synthesized to include restriction enzyme sites by GenScript U.S.A, Inc., Piscataway, New Jersey, U.S.A. Genscript then subcloned each gene into the expression plasmid pCAGGs, generously Get A Quote

摘要

The overall goal of this thesis was to determine whether envelope-free Ebola virus (EBOV) nucleocapsids could be isolated. We identified novel structures from EBOV infected cells that we have termed “proto-nucleocapsids”. These proto-nucleocapsids were ~30.5 nm in diameter and single particle image analysis revealed a hollow structure with small lobes, attributed to individual NP molecules. In addition, no large external protuberances on this structure were observed. As such, we hypothesize that the proto-nucleocapsids are the inner NP-RNA layer of the EBOV nucleocapsid. Expression of EBOV NP, VP24, VP30, VP35, and VP40 followed by density gradient

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