Cloning Of Human Developmental Control Gene-Pax2 And Its Eukaryotic Expression

OBJECTIVEThe development of mammalian embryos and organs is a much complex process; involved in a series events, proliferation of stem cells, differentiation, formation of mature cells with specific function, formation of embryo in a specific way and guide the growth of cells normally, etc. Its molecular process is much complex and stern. Genes for developmental control have a pivotal role in this process, called developmental control genes.The genes exert their affection on target cells proliferation, movement and differentiation processes through their interaction or their genetic products. Pax genes are a gene family of developmental control, that encode nuclear transcription factors and have been implicated in the control of mammalian development. As important control genes, the nuclear transcription factors are expressed in many kinds of tumor cells. Studies confirmed that Pax genes not only control the development of important organs, but they also play a key role in development and progression of malignant tumors. When adult, expression of Pax genes involve in origin of malignant tumors, act as an oncogene.Renal Cell Carcinoma (RCC) is one of the most common malignancy in the urogenital system. In late stage, there are not any effective strategy on the disease.Researches on tumor immunity found that dendritic cells (DCs) who take up variety of antigen, and express plentiful MHC molecules, costimulatory molecules and adhesion molecules, are the most powerful antigen presenting cells (APCs), and plays a pivotal role in body anti-tumor immune response. However, DCs must be pulsed with tumor antigen in order to activate the most effectively and specific anti-tumor immune response. Pulsed with polypeptide, lysate of tumor cells, apoptotic body, purified protein and nucleic acid, as vaccines, DCs have been applied to RCC patients for clinical trials.This study intend to clone Pax2 full-length open reading frame from human RCC tissues, construct an efficient eukaryotic expression recombinant vector of hPax2, generate DCs from peripheral blood PBMCs of RCC patient to prepare hPax2/DCs vaccines, then evaluate the possibility of obtaining substantial DCs which tranfected by recombinant vector with high immune activity for clinical use. METHODS...