Mouse chemokine CXCL10 eukaryotic expression vector.doc

 PAGE \* MERGEFORMAT 13 Mouse chemokine CXCL10 eukaryotic expression vector [Abstract] Application of RT-PCR method, T cells from mouse spleen mRNA, the amplified coding CXCL10 chemokine in mice of cDNA, and to add human monocyte chemoattractant protein-1 signal peptide gene was cloned to Pucm-T vector by restriction enzyme digestion and sequencing, thus building a CXCL10 recombinant eukaryotic expression vector. Liposomes in vitro using transfected COS-7 eukaryotic cells by Western blot and the chemotactic index was detected in transfected cells to express CXCL10 protein, its supernatant has a chemotactic effect on lymphocytes. CXCL10 eukaryotic expression vector was successfully constructed, in order to further study its role in tumor therapy laid the foundation. [Keywords:] CXCL10; gene cloning; eukaryotic expression vector Gene cloning and eukaryotic expression vector construction of murine CXCL10 [Abstract] The cDNA fragment encoding the murine CXCL10 was obtained from mRNA of mouse T cells by using reversal transcript-polymerase chain reaction (RT-PCR) and contained a signal peptide derived from the human monocyte chemoattractant protein 1, then it was cloned into the Pucm-T vector. By partial nucleotide sequencing, the cDNA was consistent with the reported mCXCL10 cDNA in the gene bank, and then was inserted into the eukaryotic expression vector pcDNA3.1 (). The recombinant plasmid was transfected into COS-7cell line by DOTAP liposome. The expression of the target molecule was assayed by Western blot and chemotaxis assay.It was found that the cells transfected with plasmid vector could express mCXCL10 protein and it’s culture supernatants could attract T cells markedly. The successful construction of mCXCL10 eukaryotic expression vector lays the foundation for it’s further researching in treatment of tumor. [Keywords:] CXCL10; gene cloning; eukaryotic expression vector Chemokine CXCL10 (IFN-γ inducible protein-10, IP-10), belonging to CX