2006•Unpublished venueRequires access
Expression of Recombinant Human MTIG Gene with C Terminal of His-tag in EC9706 Cells
Xinfang Houl, Qingxia Fanl, Liuxing Wangl, Ruilin Wangl, Shixin Lu
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Abstract
Objective. To construct a recombinant eukaryotic expression plasmid of MTIG and express it in human esophageal cancer cell line EC9706. Methods. The target sequence was amplified by PCR from pACT2~MTIG plasmid containing human MTIG cDNA and cloned into eukaryotic expression vector pcDNA3. l/Myc~His( - ) with C tenninal of myc epitope and 6 X His~tag. After restriction endonuclease digestion and DNA sequencing con~ finnation, the recombinant plasmid was tranfected into EC9706 cell by lipofectamine 2000. The positive monoclone was screened by G418. RT~PCR and Western blotting were used to detect the expression of mRNA and fusion pro~ tein of MTIG gene respectively. Results. The eukaryotic expression vector pcDNA3. l/Myc~His( - )~MTIG was successfully constructed and MTIG fused protein with His~tag was expressed in tranfected EC9706 cell. Conclu- sion. The human MTIG recombinant plasmid and the EC'9706 cell strain stably expressing MTIG fused protein with His~tag were obtained, which provide the basis for further study on biology functions of MTIG. (Life Science Journal. 2006;3(4):49-53J (ISSN: 1097-8135). '- . l t