Human MUC1 and GM.docVIP

 PAGE \* MERGEFORMAT 16 Human MUC1 and GM Author: Yuan, when Fang, Chang-Hong, YAN Wei Qing Yao Li Nan-Lin Ting-Wang Ling Zhang from [Keywords:] genetic engineering;, MUC1;, GM Construction and expression of eukaryotic coexpression plasmid containing human MUC1 gene and GMCSF gene [Abstract] AIM: To construct an eukaryotic coexpression plasmid containing the coding region of human MUC1 tandem repeats gene and GMCSF gene and to identify its expression in COS7 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 GMCSF gene were cloned into eukaryotic expression vector pcDNA3.1 () to construct recombinant plasmid pcDNA3.1 () MUC1GMCSF. Then the recombinant plasmid was transfected into COS7 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 GMCSF gene. The expression of MUC1 and GMCSF was detected. CONCLUSION: The suuessful construction and expression of recombinant plasmid pcDNA3.1 () MUC1GMCSF lay a foundation for further development of DNA vaccine against breast cancer. [Keywords] gene engineering; MUC1; GMCSF; coexpression vector; COS7 cell [Abstract] Objective: To construct human MUC1 tandem repeat sequence of recombinant GMCSF gene and the eukaryotic co-expression plasmid pcDNA3.1 () MUC1GMCSF, and observe the recombinant plasmid in COS7 cells. Methods: The signal peptide and coding MUC1 gene synthesis, synthetic methods such as fragment obtained by annealing sequence repeats of MUC1 gene duplication, restriction enzyme digestion and sequence analysis, and the GMCSF gene was cloned into pcDNA3.1 () eukaryotic expression vector to construct the eukaryotic co-expression plasmid pcDNA3.1 () MUC1GMCSF. To COS7 cells