efficient assembly and secretion of recombinant subviral particles of the four dengue serotypes using native prm and e proteins有效的装配和分泌重组subviral粒子的四种登革热使用本地人口、难民和移民事务局血清型和e蛋白.pdfVIP

Efficient Assembly and Secretion of Recombinant Subviral Particles of the Four Dengue Serotypes Using Native prM and E Proteins 1 1 2,4 1 1 Pei-Gang Wang *, Mateusz Kudelko , Joanne Lo , Lewis Yu Lam Siu , Kevin Tsz Hin Kwok , Martin 4 2 1 1 ´ 1,3 Sachse , John M. Nicholls , Roberto Bruzzone , Ralf M. Altmeyer , Beatrice Nal 1 Hong Kong University-Pasteur Research Centre, The University of Hong Kong, Hong Kong, China, 2 Department of Pathology, The University of Hong Kong, Hong Kong, China, 3 Department of Anatomy, The University of Hong Kong, Hong Kong, China, 4 Plate-Forme de Microscopie Ultrastructurale, Institut Pasteur, Paris, France Abstract Background: Flavivirus infected cells produce infectious virions and subviral particles, both of which are formed by the assembly of prM and E envelope proteins and are believed to undergo the same maturation process. Dengue recombinant subviral particles have been produced in cell cultures with either modified or chimeric proteins but not using the native forms of prM and E. Methodology/Principal Findings: We have used a codon optimization strategy to obtain an efficient expression of native viral proteins and production of recombinant subviral particles (RSPs) for all four dengue virus (DV) serotypes. A stable HeLa cell line expressing DV1 prME was established (HeLa-prME) and RSPs were analyzed by immunofluorescence and transmission electron microscopy. We found that E protein is mainly present in the endoplasmic reticulum (ER) where assembly of RSPs could be obser