Construction of eukaryotic expression vector pTAR-GET- DNT?R? and its expression in breast cancer COS-7 cells

Abstract

Objective: To synthesize the gene of human dominant-negative transforming growth factor beta receptor (DNTβRⅡ) by means of oligo chemic synthesis and PCR amplification, construct pTAR-GET-DNTβRⅡ eukaryotic expression vector and investigate whether its expression in breast cancer COS-7 cells. Methods:The DNTβRⅡ gene fragment was inserted into pTAR-GET to conduct a eukaryotic plasmid, then sequenced and identified by restrictive endonuclease digestion. The eukaryotic expression vector pTAR-GET-DNTβRⅡ was constructed and then transfected into COS-7 cells, to evaluate its expression by RT-PCR method. Results:The eukaryotic expression vector pTAR-GET-DNTβRⅡ was successfully constructed and could be instantaneously transfected and finally expressed in COS-7 cells. Conclusion:Full length gene of DNTβRⅡ can be synthesized by means of oligo chemic synthesis and PCR amplification. The construction of eukaryotic expression vector pTAR-GET-DNTβRⅡ and its successful expression in breast cancer COS-7 cells, may provide the basis for further study of breast cancer anti-tumor immunotherapy by adoptive immune cell therapy.