Construction of eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment gene G22 and its expression.
- Author:
Chen LUO
1
;
Xiao-juan HE
;
Yan ZHAO
;
Zhi-jie ZHANG
;
Guan-cheng LI
Author Information
1. Cancer Research Institute, Central South University, Changsha 410078, China.
- Publication Type:Journal Article
- MeSH:
Antibodies, Anti-Idiotypic;
genetics;
Antibodies, Neoplasm;
genetics;
immunology;
Eukaryotic Cells;
metabolism;
Genetic Vectors;
Humans;
Immunoglobulin Variable Region;
genetics;
Nasopharyngeal Neoplasms;
immunology;
Recombinant Proteins;
biosynthesis;
genetics
- From:
Journal of Central South University(Medical Sciences)
2008;33(1):16-20
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct a eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment (ScFv) gene G22, and to identify its expression in rectal cancer cells (CMT-93).
METHODS:The G22 gene was ligated into the sites of EcoRI and NotI of eukaryotic expression vector pcDNA3.1(+). After the identification and DNA sequencing, the recombinant plasmid pc DNA3.1(+)-G22 was stably transfected into CMT-93 cells, and the expression of G22 was detected by Western blot, flow cytometry and immunofluorescence staining.
RESULTS:Restriction enzyme digestion and DNA sequencing showed that the recombinant plasmid contained the human nasopharyngeal carcinoma anti-idiotype antibody ScFv gene G22. Transfection experiment verified that G22 gene could be expressed in CMT-93 cells in the right way.
CONCLUSION:The eukaryotic expression vector containing the human nasopharyngeal carcinoma anti-idiotype antibody ScFv gene G22 is successfully constructed and expressed, which is the basis for further study of its DNA vaccine.
