Aim: To construct an eukaryotic coexpression plasmid containing the coding region of human MUC1 tandem repeats gene and GM-CSF gene and to identify its expression in COS-7 cell.
Methods: MUC1 tandem repeats gene was obtained by synthesizing the segments. After identified by restriction endonuclease digestion analysis and DNA sequencing, MUC1 tandem repeats gene and GM-CSF gene were cloned into eukaryotic expression vector pcDNA3.1(+) to construct recombinant plasmid pcDNA3.1(+)-MUC1-GM-CSF. Then the recombinant plasmid was transfected into COS-7 cell by electroporation and the expression of target gene was detected by immunoflourescence and ELISA.
Results: Restriction analysis and DNA sequencing showed that the recombinant plasmid contained the coding region of human MUC1 tandem repeats gene and GM-CSF gene. The expression of MUC1 and GM-CSF was detected.
Conclusion: The suuessful construction and expression of recombinant plasmid pcDNA3.1(+)-MUC1-GM-CSF lay a foundation for further development of DNA vaccine against breast cancer.