Recombinant expression of Japanese encephalitis virus non-structural protein NS1 gene and its reaction with Flavivirus antigen and antibody
- Author:
ZHANG Yijia
;
YAO Xiaohui
;
CAO Lei
;
WANG Ruichen
;
FU Shihong
;
NIE Kai
;
LI Fan
;
YIN Qikai
;
HE Ying
;
WANG Huanyu
;
XU Songtao
;
MA Chaofeng
;
LIANG Guodong
- Publication Type:Journal Article
- Keywords:
Japanese encephalitis virus;
non-structural protein 1;
prokaryotic recombinant expression of protein;
antibody response to mosquito-borne flavivirus antigen
- From:
China Tropical Medicine
2023;23(12):1241-
- CountryChina
- Language:Chinese
-
Abstract:
Abstract： Objective To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.
- Full text:20240628170515320821.Recombinant expression of Japanese encephalitis virus non-structural protein NS1 gene and its reaction with Flavivirus antigen and antibody.pdf
