IL-18-PE38 eukaryotic expression plasmid and expression of.docVIP

 PAGE \* MERGEFORMAT 16 IL-18-PE38 eukaryotic expression plasmid and expression of Authors: on behalf of the Xia Lu, Li Hong, Chen Dengbang, Tsang Chun - Keywords: immunotoxins Recombinant immunotoxin is through genetic engineering technology will have a capacity-oriented molecules (carrier) and have cytotoxic molecules (toxins) coupling made of a specific cell killing ability of hybrid molecules. It has a small molecule, the product of high uniformity, high activity and heterogeneity is low, easy-to-large-scale preparation, etc., is to study the immune toxins in the process of a major leap forward. In the treatment of cancer, allograft rejection, autoimmune diseases and chronic infectious diseases has shown tremendous potential. To date, a series of recombinant immunotoxins in in vitro experiments and animal studies show good results [1 ~ 4]. In this study, first by RT-PCR received mIL-18 gene fragments, and then will have the leading role of the IL18 and have a cell killing effect of the PE38 gene construct into the IL18-PE38 fusion gene eukaryotic expression vector, and transfected into NIH3T3 cells to observe the expression of its target protein for further study of IL18-PE38 recombinant immunotoxin gene therapy of autoimmune diseases, the role of lay the foundation. 1 Materials and methods 1.1 vectors and cell lines containing the PE38 gene vector-based PRKL459K room to save. Eukaryotic expression vector Psec Tag 2B for invitrogen company’s products. Competent JM109 bacteria as a Takara product. NIH3T3 cells Cell Biology teaching and research university to provide. 1.2 Reagent RT-PCR kits for the Takara company’s products. Trizol Reagent for GibicoBRL company’s products. Restriction endonuclease, T4DNA ligase for the New England company’s products, DNAmarker for Promega products. Gel extraction kit, plasmid kit small mention of the Shanghai Waston Bio-Engineering Co., Ltd. products. DMEM medium for invitrog