Construction of a prokaryotic expression vector for apoptin and preparation of polyclonal antibody of apoptin.
- Author:
Jian-Sheng WANG
1
;
Ming-Xin ZHANG
;
Xiao-Yi DUAN
;
Fei MO
;
Quan-Ying WANG
;
Guang-Xiao YANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies; blood; genetics; immunology; Capsid Proteins; genetics; immunology; Cloning, Molecular; Escherichia coli; metabolism; Gene Expression; Genetic Vectors; Genome; Mice; Mice, Inbred BALB C; Plasmids
- From: Journal of Southern Medical University 2009;29(7):1405-1407
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a prokaryotic expression vector for apoptin and prepare polyclonal antibody of apoptin.
METHODSApoptin gene amplified from pGEM-T/Apoptin plasmid by PCR was cloned into pET-28a (+). E.coli BL21 (DE3) was transformed by the recombinant plasmid, and apoptin protein expression induced by IPTG was analyzed by SDS-PAGE. BALB/c mice were immunized with the protein and the titer of the antibody was determined using indirect enzyme-linked immunosorbent assay (ELISA).
RESULTSApoptin gene was successfully cloned into pET-28a (+), and the expression of a protein with relative molecular mass of about 17 000 was identified by SDA-PAGE. After 5 immunizations of the mice with the protein, the blood antibody titer reached 1:5x10(5).
CONCLUSIONThe prokaryotic expression vector for apoptin is successfully constructed and the polyclonal antibody of apoptin is obtained, which allows further functional study of apoptin.
