Construction and expression of eukaryotic coexpression plasmid containing human MUC1 gene and GMª²CSF gene

    YUAN Shiª²fang1, SHI Changª²hong2, YAN Wei3, YAO Qing1, LI Nanª²lin1, WANG Ting1, WANG Ling1, ZHANG Yingª²qi4

    1Department of Vascular and Endocrine Surgery, Xijing Hospital; 2LaboratoryAnimalCenter,3DepartmentofPathology,4Biotechnology Center, Fourth Military Medical University, Xi¡¯an 710032, China

    \[Abstract\] AIM: To construct an eukaryotic coexpression plasmid containing the coding region of human MUC1 tandem repeats gene and GMª²CSF gene and to identify its expression in COSª²7 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 GMª²CSF gene were cloned into eukaryotic expression vector pcDNA3.1(+) to construct recombinant plasmid pcDNA3.1(+)ª²MUC1ª²GMª²CSF. Then the recombinant plasmid was transfected into COSª²7 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 GMª²CSF gene. The expression of MUC1 and GMª²CSF was detected. CONCLUSION: The suuessful construction and expression of recombinant plasmid pcDNA3.1(+)ª²MUC1ª²GMª²CSF lay a foundation for further development of DNA vaccine against breast cancer. ......