Constructions Of Reporter Genes' (lux, GFP) Vectors And Studies Of Their Expressions In Tumor Cells And Tissues

Tumor is a fatal disease to human health, the key problem to tumor diagnosis and biological (gene) therapy is the targetting and specific location of the Tumors, one of the efficient ways for solving the problem is the use of both reporter gene and tissue and cell specific promoters and enhencers. Among the commonly used reporter genes, bacterial bioluminesent gene lux and Green fluorescent protein are the two new and promising reporter genes in consequence of their no harm to cells and easy detection. Till now there are no reports of the lux genes especially by the use of mammalian expression vectors pcDNA₃ and under the direction of AFP and HBV, making lux and GFP express in human tumor cells and tissues. In order to study the possibilities of the expressions of lux genes in various tumor cells, compare the characteristics of the expression and availability of lux and GFP reporter genes in tumor cells and tissues, investigate the utilities of the specific expressions of luxAB and GFP genes under the direction of specific promoters and enhencers, and provide the basis for the use of reporter genes especially the lux gene in the study of tumor metastasis, diagnosis and specific targetting gene therapy, Fused bacterial luciferase luxAB and GFP gene mammalian expression vectors were constructed by site directed mutagenesis and genetic engineering and expressed in tumor cells by lipofectin transfection , serial luxAB and GFP gene mammalian expression vectors under the direction of AFP or HBV were also constructed and expressed (or specific) in different tumor cells, at last, expressed luxAB and GFP transfected Tumor cells were stably injected into nude mice subcutaneously for the study of their...