Endothelin-1 series bodies in COS7 Construction and expression of cell identification.docVIP

 PAGE \* MERGEFORMAT 7 Endothelin-1 series bodies in COS7 Construction and expression of cell identification [Abstract] Objective: Cloning and Expression of endothelin-1 (endothlin1, ET1) series body, to improve the immunogenicity of ET1 resulting from the body and its immune antibody titer. Methods: pGEX  4T  1 vector of ET1 genes of the series, and the ET1 gene downstream of the introduction of tandem GM  CSF gene, restriction enzyme digestion and sequence analysis of the recombinant plasmid; then ET1 string containing the body and GM  CSF fusion gene connected to the eukaryotic expression vector pcDNA3.1 () in the , the same after restriction enzyme digestion and sequencing confirmed; and to construct expression vector was transfected into COS  7 cells by indirect immunofluorescence assay and ELISA assay showed that gene in COS  7 cells. Results: Correct construction of the ET1 Repeat four strings GM  CSF gene and cloning of recombinant vector pGEX  ET1 ④  GM  CSF, as well as the eukaryotic co-expression plasmid pcDNA3.1  ET1 ④  GM  CSF; indirect immunofluorescence assay and ELISA analysis showed that gene ET1 and GM  CSF in COS  7 cells have been expressed. Conclusion: Endothelin-1 series bodies Construction and COS  7 cells will be successful expression of the immunological characteristics of ET1, nucleic acid vaccine and related diseases, self - Immunotherapy lay the foundation for further study. [Keywords:] endothelin-1 COS  7 cells in autoimmune 0 Introduction Endothelin-1 (endothelin1, ET1) in 1988 by Yanagisawa et al [1-2] from the culture supernatant of porcine aortic endothelial cells were isolated and purified a small molecule vasoconstrictor peptide, widely distributed in the nervous system, circulation system and the endocrine system, with a wide range of biological activity, as a potent vasoconstrictor peptide and vascular smooth muscle cell mitogen involved in myocardial ischemia, hypertension, stroke,