Human GM-CSF gene eukaryotic expression vector and expression of.docVIP

 PAGE \* MERGEFORMAT 12 Human GM-CSF gene eukaryotic expression vector and expression of 【Abstract】 【Objective】 To construct gene adjuvant hGM-CSF gene-based eukaryotic expression plasmid pIGM-CSF, and expression in cervical carcinoma HeLa cells, and identification. 【Methods】 polymerase chain reaction from the carrier pORF-hGM-CSF in hGM-CSF gene was amplified, cloned into the bicistronic eukaryotic expression vector pIRES of the multiple cloning site B (MCSB), the construct pIGM - CSF eukaryotic expression vector. Then using Lipofectamine law will HeLa cells transfected using RT-PCR in RNA level and the ELISA at the protein level was detected in eukaryotic cells. 【Results】 The cloned hGM-CSF gene fragments by DNA sequencing entirely correct; recombinant plasmid transfected Hela cells, tests, hGM-CSF expression. 【Conclusion】 containing gene was successfully constructed the eukaryotic expression vector adjuvant pIGM-CSF, and to be effective in cervical cancer cells express antigens for the purpose of cloning in order to continue to construct bicistronic eukaryotic expression vector basis. Keywords: eukaryotic expression vector; hGM-CSF; gene adjuvant; bicistronic Abstract: 【Objective】 To construct eukaryotic expression plasmid pIGM-CSF based on gene adjuvant hGM-CSF, and express in Hela cells. 【Methods】 Plasmid pORF-hGM-CSF was used as template to amplify hGM-CSF gene by PCR, and the product was inserted into multiple cloning site’s B of bicstronic eukaryotic expression vector pIRES to construct eukaryotic expression plasmid pIGM-CSF, then transfected into Hela cell by using liposome. Finally, the hGM-CSF gene expression of protein was detected by RT-PCR from RNA level and ELISA from protein level. 【Results】 The length and sequence of the cloned hGM-CSF segment was correct. The hGM-CSF gene could be expressed in transfected Hela cells. 【Conclusion】 The eukaryotic expression plasmid pIGM-CSF containing gene adjuvant was successfully constr