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Updated: Jul 17, 2026
A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells
Published on: June 10, 2015
[Construction and expression of eukaryotic coexpression plasmid containing human MUC1 gene and GM-CSF gene]
Shi-fang Yuan1, Chang-hong Shi, Wei Yan
1Department of Vascular and Endocrine Surgery, Xijing Hospital, China. shifangy@fmmu.edu.cn
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.
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