Generation of high titer antisera of a novel liver-specific gene FF456 in mice by DNA immunization and it’s application
- VernacularTitle:DNA免疫制备肝特异新基因FF456抗体及其应用
- Author:
Guangwu HE
;
Chang SHU
;
Hao SHEN
;
Minyue ZHANG
- Publication Type:Journal Article
- Keywords:
FF456;
DNA immunization;
Antisera;
Immunofluorescence
- From:
Journal of Medical Postgraduates
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: FF456 is a new gene cloned in our lab, which belongs to c-type G protein-coupled receptors and has high homology to the olfactory receptor family. Northern blotting and RT-PCR showed that the expression of FF456 was exclusively restricted in liver and was significantly down-regulated in hepatoma. In an effort to study the function of FF456 gene, high titer antibody is indispensable. Methods:Full-length cDNA of FF456 was reconstructed into pcDNA3.1(-) vector, which was used for immunizing 6- to 8- weeks old female BALB/c mice. The effects of different injection manners, solvents and dosages on the antibody production have been compared. For detecting the titer and specificity of the antisera produced by DNA immunization, a peptide containing FF456 specific sequence was expressed by E.coli expression system. Results:Finally specific antisera with titers as high as 1 ∶ 50 000 was obtained. Conclusion:Immunofluorescence assays showed FF456 was expressed in hepatocytes with high tissue specificity. The FF456 expression in hepatoma cells was decreased dramatically.
