Gene optimization, protein expression, purification and characterization of the HDV antigen for diagnosis.
- Author:
Jun-Ying DING
1
;
Min-Zhuo GUO
;
Yao YI
;
Qiu-Dong SU
;
Xin Lu XUE
;
Feng QIU
;
Sheng-Li BI
Author Information
- Publication Type:Journal Article
- MeSH: Enzyme-Linked Immunosorbent Assay; Hepatitis D; diagnosis; Hepatitis delta Antigens; genetics; immunology; isolation & purification; Humans; Plasmids; Recombinant Proteins; biosynthesis; immunology; isolation & purification
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(2):87-89
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare HDAg with biological activities as a candidate of diagnostic reagent.
METHODSTo synthesize HDV gene fragment after codon optimization. To construct a thio-fused recombinant plasmid based on M48 expression vector. To express in E. coli induced by IPTG. To purify the protein by affinity chromatography followed by characterization in ELISA:
RESULTSPlasmid construction was verified by enzyme digestion. SDS-PAGE indicated the molecular weight of the protein was the same as we expectation. ELISA proved its affinity with HDV antibodies.
CONCLUSIONHDAg was obtained successfully and it will pave the road to the research of HDV diagnostic reagent.
