Construction of pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? eukaryotic expression vector and expression in NIH 3T3 cells
- VernacularTitle:真核表达载体pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/?的构建及其在NIH 3T3细胞株中的表达
- Author:
Yongxian HU
;
Kang YU
;
Yingxia TAN
;
Jianbo WU
;
Zhijian SHEN
;
Honglan QIAN
;
Bin LIANG
;
Daming SHAN
- Publication Type:Journal Article
- Keywords:
Antibodies;
Neoplasms;
Gene therapy;
Gene expression;
NIH 3T3 cells
- From:
Chinese Journal of Pathophysiology
1999;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To construct a recombinant eukaryotic expression vector pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? and detect its expression in NIH 3T3 cells.METHODS:CD28-? cDNA was amplified from the plasmids pBULLET and inserted into pLNCX vector that contained anti-CD20 scFv/IgGFc/CD80 gene.The recombinant plasmids were transfected into NIH 3T3 cells,and resistant clones were obtained by G418 selection.The gene expression of the fusion protein was determined by RT-PCR and FACS.RESULTS:The recombinant eukaryotic vector was constructed successfully,determined by PCR and enzyme digestion analysis.The target gene was amplified from NIH 3T3 cells transfected with the vectors by RT-PCR.The FACS showed that recombinant protein was expressed in NIH 3T3 cells.CONCLUSION:Construction of pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? expression vector and its expression in NIH 3T3 cells lay the foundation for further research of generation of modified T lymphocytes to CD20 positive lymphoma.
