登革病毒NS1蛋白在Sf9-杆状病毒系统中的表达与纯化

Expression and Purification of Dengue NS1 Protein in Sf9-Baculovirus System.

作者信息

Coelho Diego R, Carneiro Pedro Henrique

机构信息

Department of Medicine, Feinberg School of Medicine and the Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, IL, USA.

Laboratory of Structural Biotechnology and Bioengineering, Biophysics Institute Carlos Chagas Filho, Lab C0-36ss, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

出版信息

Methods Mol Biol. 2022;2409:39-46. doi: 10.1007/978-1-0716-1879-0_4.

DOI:10.1007/978-1-0716-1879-0_4

PMID:34709634

Abstract

It is well known that glycosylations of Dengue NS1 protein are important for its structure, oligomerization, and immunogenicity. One of the major challenges in heterologous NS1 protein expression is the difference in glycosylation patterns amongst different organisms. The two major natural hosts for Dengue virus are humans and mosquitoes, which are capable of producing very complex glycosylation motifs. This chapter presents an optimized protocol for heterologous expression and purification of Dengue NS1 protein in Sf9 cells infected with baculovirus. NS1 protein obtained from this protocol is glycosylated and capable of forming soluble hexamers that can be used for structural and functional assays.

摘要

众所周知，登革热NS1蛋白的糖基化对其结构、寡聚化和免疫原性很重要。异源NS1蛋白表达的主要挑战之一是不同生物体之间糖基化模式的差异。登革热病毒的两个主要天然宿主是人类和蚊子，它们能够产生非常复杂的糖基化基序。本章介绍了一种在感染杆状病毒的Sf9细胞中异源表达和纯化登革热NS1蛋白的优化方案。通过该方案获得的NS1蛋白被糖基化，能够形成可用于结构和功能分析的可溶性六聚体。