Study On The Mechanism Of Metabolic Abnormalities In Large - Capacity Transgenic And Otg1 Mutations Of PiggyBac Transposon

The ability to reliably introduce individual full-length genes into the mammalian and human genomes is essential for transgenic studies and gene therapy. However, the small transgene size, the uncontrollable gene copy number, and the lack of defined transgene integration orientation of the current gene transfer systems represent a major hurdle in the application of transgenesis and gene therapy. We showed that the modified piggyBac (PB) transposon system can efficiently introduce an intact mammalian gene of more than200kb into human and mouse cells and in mice. PB-mediated transgenesis permits the stable integration and inheritance of single-copy full-length mammalian genes, and allows reliable gene expression that mimics the endogenous ones. The modified PB transposon system facilities the stable integration and inheritance of large DNA fragments, and allows controlled expression of the transgene. The vector has the capacity to carry regulatory elements associated with a gene and the positional effects are not obvious. PB could serve as an effective and reliable high capacity vector for functional analysis of the mammalian genome and for gene therapy in human cells.The precise regulation of glucose metabolism is the basic guarantee for the body to perform normal physiological function. Insulin resistance and diabetes with the main feature of abnormally high blood glucose levels are important health challenges for modern society, and low blood glucose will cause vertigo、convulsions and other adverse reactions. Serious low blood will cause damage to the brain and is life-threatening. We obtained a mutant line for novel gene Otgl during PB transposon mediated mutagenesis screen in mice, and we found that Otgl protein locate in the Golgi. Otgl mutant mice exhibited hypoglycemia、hypoinsulinemia、growth retardation and died after birth, these data indicate that Otgl protein is involved in glucose metabolism. This study further showed that Otgl mutant mice also showed decreased growth hormone and IGF-1, less insulin secretory vesicles. We obtained many candidates including CaMKⅡ which are predicted to interact with Otgl through mass spectrometry, and we found decreased phospho-CaMKⅡ level in Otgl mutant mice, indicating that Otgl may regulate hormone secretion through CaMKⅡ. The study of Otgl gene function may provide some clues for the mechanism and signal pathway research of human perinatal mortality disease caused by metabolism disorder, also our study will give some perspectives for glucose metabolism and protein trafficking.