Construction of eukaryotic expression vector encoding human nasopharyngeal carcinoma anti-idiotype antibody single chain fragment gene G22 and its expression / 中南大学学报(医学版)

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.