[Construction of eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment gene G22 and its expression].
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.


CONCLUSIONS
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.
Luo
Chen
C
Cancer Research Institute, Central South University, Changsha 410078, China.
He
Xiao-juan
XJ
Zhao
Yan
Y
Zhang
Zhi-jie
ZJ
Li
Guan-cheng
GC
D000888:Antibodies, Anti-Idiotypic / Q000235:genetics*; D000912:Antibodies, Neoplasm / Q000235:genetics / Q000276:immunology*; D005057:Eukaryotic Cells / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics; D009303:Nasopharyngeal Neoplasms / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D000888:Antibodies, Anti-Idiotypic; D000912:Antibodies, Neoplasm; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins