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Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography.


ABSTRACT: The norovirus capsid is composed of a single major structural protein, termed VP1. VP1 is subdivided into a shell (S) domain and a protruding (P) domain. The S domain forms a contiguous scaffold around the viral RNA, whereas the P domain forms viral spikes on the S domain and contains determinants for antigenicity and host-cell interactions. The P domain binds carbohydrate structures, i.e., histo-blood group antigens, which are thought to be important for norovirus infections. In this protocol, we describe a method for producing high quality norovirus P domains in high yields. These proteins can then be used for X-ray crystallography and ELISA in order to study antigenicity and host-cell interactions. The P domain is firstly cloned into an expression vector and then expressed in bacteria.

SUBMITTER: Leuthold MM 

PROVIDER: S-EPMC4941939 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature